Many people with chronic fatigue syndrome are infected with a little known virus that may cause or at least contribute to their illness, researchers are reporting. 

The syndrome, which causes prolonged and severe fatigue, body aches and other symptoms, has long been a mystery ailment, and patients have sometimes been suspected of malingering or having psychiatric problems rather than genuine physical ones. Worldwide, 17 million people have the syndrome, including at least one million Americans. 

An article published online Thursday in the journal Science reports that 68 of 101 patients with the syndrome, or 67 percent, were infected with an infectious virus, xenotropic murine leukemia virus-related virus, or XMRV. By contrast, only 3.7 percent of 218 healthy people were infected. Continuing work after the paper was published has found the virus in nearly 98 percent of about 300 patients with the syndrome, said Dr. Judy A. Mikovits, the lead author of the paper.

XMRV is a retrovirus, a member of the same family of viruses as the AIDS virus. These viruses carry their genetic information in RNA rather than DNA, and they insert themselves into their hosts’ genetic material and stay for life. 

Dr. Mikovits and other scientists cautioned that they had not yet proved that the virus causes the syndrome. In theory, people with the syndrome may have some other, underlying health problem that makes them prone to being infected by the virus, which could be just a bystander. More studies are needed to explain the connection. 

But Dr. Mikovits said she thought the virus would turn out to be the cause, not just of chronic fatigue, but of other illnesses as well. Previous studies have found it in cells taken from prostate cancers. 

“I think this establishes what had always been considered a psychiatric disease as an infectious disease,” said Dr. Mikovits, who is research director at the Whittemore Peterson Institute in Reno, a nonprofit center created by the parents of a woman who has a severe case of the syndrome. Her co-authors include scientists from the National Cancer Institute and the Cleveland Clinic.

Dr. Mikovits said she and her colleagues were drawing up plans to test antiretroviral drugs — some of the same ones used to treat HIV infection — to see whether they could help patients with chronic fatigue. If the drugs work, that will help prove that the virus is causing the illness. She said patients and doctors should wait for the studies to be finished before trying the drugs.

Dr. William Schaffner, an infectious disease expert at Vanderbilt University, said the discovery was exciting and made sense. 

“My first reaction is, ‘At last,’ ” Dr. Schaffner said. “In interacting with patients with chronic fatigue syndrome, you get the distinct impression that there’s got to be something there.”

He said the illness is intensely frustrating to doctors because it is not understood, there is no effective treatment and many patients are sick for a long time. 

He added, “This is going to create an avalanche of subsequent studies.”

To many people who suffer from the poorly understood illness called chronic fatigue syndrome, a recent study linking the disorder to a virus was a victory for the little guys.

For one thing, the study pointed to a physical cause for an illness that the medical establishment had often snidely dismissed as psychosomatic. The research could not be ignored: it was published last month in Science, one of the world’s pickiest and most prestigious journals. The discovery came, in a sense, from within the patients’ own ranks: several of the scientists, including the lead author of the report, worked for the Whittemore Peterson Institute for Neuro-Immune Disease, a nonprofit in Reno, Nev., founded recently by the parents of a young woman who has the syndrome. And even though the institute was new, it had attracted collaborators from two high-powered centers, the National Cancer Institute and the Cleveland Clinic. 

Harvey and Annette Whittemore were not the first to start a research foundation out of desperation to find answers for an incurable disease. The actor Michael J. Fox did the same for his own illness, Parkinson’s disease. Others have created groups for pancreatic cancer, amyotrophic lateral sclerosis (Lou Gehrig’s disease) and breast cancer. The foundations have in common a desire to pick up the pace of research, often by financing innovative ideas and avoiding red tape and bureaucracy. But few if any of the private groups have produced notable results as quickly as the Whittemore Peterson Institute has. 

At least one million Americans have chronic fatigue syndrome, which causes severe fatigue, muscle and joint pain, sleep problems, difficulty concentrating and other problems. Its cause is unknown, symptoms can last for years and there is no effective treatment. Women outnumber men as patients, and many people with the syndrome feel stigmatized and brushed off as neurotic by doctors. Andrea Whittemore-Goad, who is 31, has had a severe case of the syndrome for 20 years. 

“Three major medical institutions tried to marginalize her, tell her she didn’t want to get well,” Annette Whittemore said in an interview. 

But Mrs. Whittemore had seen her daughter turn abruptly from a happy, healthy child to a very sick one, and she has steadfastly believed that some sort of infection must have been the cause. Researchers have not pursued the infection theory seriously enough, in her opinion, and doctors have been scornful. One physician told her that if he could not understand her daughter’s lab results, they couldn’t be important.

Another said “he didn’t look at my daughter’s medical information because he didn’t want the facts to get in the way of his theory,” Mrs. Whittemore said in an e-mail message. “I could write a book on ridiculous things that doctors say to patients with C.F.S.” She added: “One day I just had enough. I said, ‘I’ve got to do something.’ ”

She and her husband had the means, the knowledge and the connections. They are real estate developers and part owners of a gas company and an energy drink, and they hold interests in other businesses. Mr. Whittemore is also a lawyer and a lobbyist. Starting in the fall of 2004, they put $5 million of their own money into setting up an institute at the University of Nevada’s medical school. They also persuaded the governor and State Legislature to commit $10 million for a new building that would house the institute’s researchers and a clinic, as well as scientists from the university and the Nevada Cancer Institute. The research began in 2006, and a clinic for patients is scheduled to open in about a year.

Rather than just doling out money to far-flung researchers, the Whittemores wanted to employ their own scientists who would be devoted full time to the cause. In the spring of 2006, they met Dr. Judy A. Mikovits, a virus expert who had spent 22 years working at the National Cancer Institute. She had left the institute in 2001 to get married and move to California, where she went to work for a drug development company that failed. She was tending bar at a yacht club when a patron said her constant talk about viruses reminded him of someone he knew in Nevada. That person was a friend of Annette Whittemore’s. Dr. Mikovits soon found herself at a conference on chronic fatigue syndrome.

“The meeting was terrible,” she said. “The science in C.F.S. is awful, not because the researchers are awful, but because they have no resources.” 

But one presentation made her practically leap out of her seat. The speaker was Dr. Daniel L. Peterson, who treats Ms. Whittemore-Goad (he is the Peterson for whom the institute is named). He has seen about 5,000 patients with the syndrome in the last 20 years, and he described some who had also developed a rare type of lymphoma. 

“I said, ‘That’s a retrovirus,’ ” Dr. Mikovits recalled. 

Dr. Mikovits began connecting the dots almost immediately. She knew that some patients with chronic fatigue syndrome, and some men with prostate cancer, had a certain enzyme deficiency. And she also knew that tissue samples from men with prostate cancer had been found to harbor a retrovirus called XMRV, for xenotropic murine leukemia virus-related virus. She began working part time with the institute, and by the fall of 2006, the Whittemores had hired her as research director. One of her first projects was to look for XMRV in blood samples from people with chronic fatigue syndrome and from healthy control subjects. 

Many of the samples from syndrome patients — 68 of 101, or 67 percent — were infected, she and her colleagues reported in Science. Only 3.7 percent of the healthy controls carried the virus. XMRV, the scientists suggested, may cause or at least contribute to chronic fatigue syndrome. Further tests found the virus in 90 of the 101, Dr. Mikovits said.

Retroviruses can cause cancers in animals, and in humans, they include HIV and a virus that can cause leukemia. “I knew how serious a retrovirus is,” Mrs. Whittemore said. “I was very concerned, knowing there would be serious implications. My second thought was, of course it was going to be something serious like that. Look at my daughter and how ill she is. Why would we expect it to be something simple? I also felt like the weight of world was on my shoulders. We would have to be telling people some very bad news.”

But many patients were grateful, and said they found hope in the discovery — hope that now their illness would be taken seriously, and hope that the new information would lead to effective treatments or even a cure. 

But the study is not conclusive. Other scientists need to replicate the tests to find out if the initial findings hold up, and a great deal more work is needed to determine whether XMRV really does play a role. Just detecting it in patients does not prove it made them sick; patients may have some other underlying problem that makes them susceptible to XMRV, which could be just a passenger in their cells.

Dr. Peterson said he thought the crux of the problem in chronic fatigue syndrome involved a derangement in the immune system that could allow old viral infections — with various herpes viruses, for instance — that were once under control to become active again.

“XMRV is the sort of agent that could create that effect on the immune system,” Dr. Peterson said. But he emphasized that more research was needed.

AMONG those expected to try to replicate the XMRV findings is the Centers for Disease Control and Prevention. But Dr. [Dr. William Carlisle Reeves (born 1943)], who directs the agency’s research on the syndrome, has said that he does not expect to find the virus in blood samples from patients. He said that no other studies had ever proved a virus to be the cause, and that stress and a history of sexual and emotional abuse were more likely to play a role in many cases. “I blame the C.D.C. for most of this mess,” Mrs. Whittemore said, adding that thinking like Dr. Reeves’s was what drove her to start her own research center.

The Whittemore Peterson Institute is running on about $1 million a year, to pay for salaries, equipment, supplies and other expenses. In the world of research, that is not much. The Whittemores are trying to raise more money, and have received donations as large as $50,000 from friend down to a few dollars from patients who are sick, out of work and broke, but eager to support any research that may one day help them.

Mrs. Whittemore feels that she is racing against time to save her daughter. For a while, Ms. Whittemore-Goad had improved on an antiviral drug, but she had to stop taking it because she had a reaction to it. She had worked part time as a yoga instructor, and she married in the last year, but now rarely goes out because she has seizures.

“She is just really very, very sick,” Mrs. Whittemore said. “She’s had this for a long time. We’ve got to get something for her as soon as possible. That is the driver for the speed on this. I don’t want to lose her. I can’t lose my daughter. I don’t want to win this battle and lose the war.” '

The Patient Advocate took a picture of Judy Mikovits from the 2009 UK conference DVD and gave it to his daughter, saying, “This is the woman who is going to make you well”. My daughter put Judy’s picture on the wall of her room where she rests.

The Patient Advocate first saw Judy Mikovits at the CFS conference in Ft. Lauderdale, FL in January 2007. She was presenting a paper on “Incidence of Chromosomally Integrated HHV-6 (CIHHV-6) in a Cohort of CFS patients with Clonal TCR-g and Lymphoid Malignancies”. The PA did not take any special note of this lecture except to note that Dr. Mikovits talked like a typewriter.

In April of 2008, the PA saw Dr. Mikovits give another presentation in London. The PA’s report at the time said this: “The last major lecture was an incredibly high-powered presentation by the American researcher, Dr. Judy Mikovits. She is the research director of the new Whittemore-Peterson Institute of Neuro-Immune disease in Reno, NV. In a lecture of which I understood next to nothing, she gave every indication that this institute has the funding, the drive and the independence to reveal some important elements of this disease. She also indicated a no-nonsense willingness to cooperate with others world-wide in this struggle. I have seen many scientists make presentations, and this gal was amazing. With this presentation, along with the others, there was a clear picture that the Americans were back in the UK doing what they do best.”

Another attendee of this conference wrote this: “Dr Judy Mikovits is Research Director of the Whittemore Peterson Institute in Nevada and she presented a most enthralling presentation on the focus the Institute's research although most of the technical detail was way over my head. The research looks at abnormal inflammatory response to viruses and part of it piggybacks off Dr Kerr's work.”

Of the Reno conference in 2009, the PA wrote this: Mikovits, the director of the Whittemore-Peterson gave a slam-bang talk, one similar to what she delivered in the UK last May. However, this time she had less time and she really had to whirl through it. This gal is generally accepted as being a serious researcher and in a position to get some results. She is obviously bringing money and people over from her former research field in cancer. Here is a quote from Judy Mikovits that surfaced on Cort Johnson’s site after the Reno conference: “We have a piece of data that would just knock your socks off but I’m not showing it. “

The PA has developed the habit of sitting through days of scientific or quasi- scientific lectures. It is possible to nose out the more scientifically interesting talks, even for a non-scientist. After awhile one can sense the “cannon fodder” talks that dominate all of these conferences – those happy little academic talks that are going to go nowhere.

Consequently this Patient Advocate was ready to really listen to Judy Mikovits in May 2009 at the London CFS/ME conference. “Dr. Judy Mikovits, who astonished so many folks at this conference last year, was back for another lecture. She made her usual complex and far reaching talk, most of which passed over my head. (I should have studied biochemistry.) Her work at the Institute involves sub-setting CFS through gene array chips and cytokine testing. The Institute is moving very fast and I get a sense that specific information is going to emerge here sooner rather than later. Certainly talking to Annette Whittemore and hearing Mikovits and Peterson makes one sense something is afoot, although they are smart enough not to say anything. During the question period Dr. Mikovits was asked if they had been working on a specific virus involved in CFS. She coyly answered, almost in a whisper, “yes” - and then she followed up that there is a novel virus not previously associated with CFS and that the WPI had submitted their research paper for publication at Science magazine.”

And then came October 9, 2009 and the world of CFS/ME turned.

The connection between Judy Mikovits and the Whittemore-Peterson Institute was a bit of serendipity. Dan Peterson was giving the last lecture of the day at a HHV6 conference in Spain in 2005. Judy Mikovits was in the audience and responded to Dr. Peterson’s plea to the audience (during the question period) for help with a problem that he was having. Ten of his CFS patients had developed mantle cell lymphoma and he was stumped. The following comes from an interview with Mikovits on Cort Johnson’s site: “These people were getting lymphoma and they were getting it before anyone gets lymphoma and to me that says virus.” “My background is in virus caused cancers. My training for over 22 years at the NCI was in how viruses dysregulate the immune system and cause disease – cancer. So when he told me to come meet with him after the talk I beat my way up to the front of the room because it was the only interesting thing that I had heard at the meeting, and I said, “There is something there, that’s a smoking gun”, so I gave him my card, told him that I’m interested, told him what I thought was going on, and he and Annette invited me out to Reno. Within two weeks of that meeting, they talked about setting up a summer research program.”

Judy Mikovits showed up in the right place at the right time. However, she is not solving this problem on her own. Many physicians and researchers have plowed the ground ahead of her. She is the beneficiary of many efforts.. It is a bit like volleyball. The ball is controlled, set up and then spiked. Judy is the spike.

From all accounts, Dr. Mikovits has great empathy with the CFS/ME patient population. All good artists have empathy for their subject. An artist, as well as a scientist, benefits from “attachment to the subject”. This empathy, this connection. does not come easy, and is not taught in schools. Not everyone has it. It is inherent in the personality of the individual. Dr. Mikovits seems to “have it”. A recent article in the Reno Gazette Journal had a personal bit about Dr. Mikovits: "My grandfather was dying of cancer, and nobody would talk about it," Mikovits said. "One day I said to him, 'Don't you know you're dying?' And he said, 'Don't say that. I don't want your grandmother or your dog, George, to know." "He was dying of cancer, and nobody would talk about it. That's what bothered me most."

Annette Whittemore has spoken of how much Dr. Judy Mikovits lab collaborators enjoy working with her. “Dr. Judy”, as her students call her, is both disciplined and relaxed, with a good sense of humor. “Dr. Judy” works long hours, setting the tone and direction. Others follow suit.

A recent article in the Reno Gazette Journal has a number of good quotes from Mikovits.

About the moment of truth with XMRV: “It was January 22, and we were in a San Diego restaurant called the Yard House,” said Mikovits, who had gone there with fellow scientist Vincent Lombardi to present the results of their research to [Dr. Francis William Ruscetti (born 1943)] and Robert Silverman, two of the world’s leading virologists. “We kept waiting for them to say something,” Mikovits said. “I was nauseous. Bob (Silverman) waited a long a time, and then he looked up and said, ‘Well, this is going to change their world.’”  And it has.

About getting traction: “They want us to send them the (XMRV) retrovirus so they can screen huge libraries of compounds and see what they have that could work,” Mikovits said. “They already have drugs to treat HIV, so they could redesign a compound for the XMRV virus. Since they already have FDA approval, they could get something out to people fast.”

"I've gotten more than 100 e-mails from physicians, and I've gotten calls from the head of the Mayo Clinic and the clinical director at Sloan-Kettering," she said. "Those people wouldn't take the time if they didn't think this was a significant finding."

"We've got about 500 samples from the United Kingdom, and we also were approached by the head of the syndrome organization in Spain," she said. "We eventually expect to collaborate with many other countries: Norway, Germany and others. Everybody is asking if we can test their samples and asking how they can get in the clinical trials."

About the Barcelona presentation in October 2009: "They applauded before I gave my presentation," she said. "It was such a surprise, because they never applaud before a presentation."

Posted by consuegra at 7:28 PM 1 comment: 

Dr. Mikovits obtained her Ph.D. in Biochemistry and Molecular Biology from George Washington University. 

Dr. Mikovits spent more than 20 years at the National Cancer Institute in Frederick MD  where she investigated mechanisms by which retroviruses dysregulate the delicate balance of cytokines in the immune response. 

This work led to the discovery of the role aberrant DNA methylation plays in the pathogenesis of HIV. Later in her career at the NCI, Dr. Mikovits directed the Lab of Antiviral Drug Mechanisms (LADM) a section of the NCI's Screening Technologies Branch in the Developmental Therapeutics Program. The LADM's mission was to identify, characterize and validate molecular targets and to develop high-throughput cell-based, genomic and epigenomic screens for the development of novel therapeutic agents for AIDS and AIDS-associated malignancies (Kaposi's sarcoma). 

Dr. Mikovits served as a senior scientist at Biosource International, where she led the development of proteomic assays for the Luminex platform that is used extensively for cytokine activity assessment in therapy development. 

She also served as Chief Scientific Officer and VP Drug Discovery at Epigenx Biosciences, where she lead the development and commercialization of DNA methylation inhibitors for cancer therapy and of cell and array-based methylation assays for drug discovery and diagnostic development.   [See EpiGenX Pharmaceuticals ]

She is Research Director at the Whittemore Peterson Nevada CFS centre for Neuro-Immune disorders and has co-authored over 40 peer reviewed publications that address fundamental issues of viral pathogenesis, hematopoiesis and cytokine biology.

Formally trained as a cell biologist, molecular biologist and virologist, Dr. Mikovits has studied the immune response to retroviruses and herpes viruses including HIV, SIV, HTLVI, HERV, HHV6 and HHV8 with a special emphasis on virus host cell interactions in cells of the hematopoietic system including hematopoietic stem cells (HSC). 

Dr Mikovits was co-author of the "Detection of an Infectious Retrovirus, XMRV, in Blood Cells of Patients with Chronic Fatigue Syndrome" research paper in October 2009 in Science magazine - click here

(with grateful thanks to the Whittemore-Peterson Institute for this information)

Long-time ProHealth subscriber and contributor Sandy Miarecki has compiled and edited in one transcript the XMRV Seminar that Whittemore Peterson Institute Research Director Dr. Judy Mikovits presented Jan 22 in Santa Barbara – helpfully indicating where each slide fits in the text. To download the slides (about 19 MB):  click here.

__________________

• Annette Whittemore: 
  • • I am happy that the ProHealth organization was able to get this online so a lot of patients who are too ill to make it are able to follow online. I’d like to thank Rich Carson, I’d like to thank ProHealth for putting this together, and [The HHV-6 Foundation] and Kristin Loomis. It’s a pleasure to be here and to have an opportunity to speak to you about the recent discovery of XMRV in Chronic Fatigue Syndrome patients.  Thank you for inviting Judy Mikovits today.  We’ve made a special effort to learn more about the most exciting news in the world of CFS since the 1980s when major outbreaks of this disease were reported in several locations around the US, including three small towns in Nevada.
    • These reports came on the heels of the discovery of HIV and AIDS.  During this time, Dr Mikovits was at the NCI working in laboratories that were actively studying this new virus, when she began her doctoral program at George Washington University.  In fact, she wrote and presented her doctoral thesis on HIV latency, presenting it the same day that Magic Johnson announced he was HIV positive.  The good news is that he is still healthy after all these years, having had the opportunity perhaps to prevent a high viral load from ever occurring.
    • Jumping forward to 1989 brings a critical event to the life of our family.  Our daughter became ill, and suddenly we found ourselves in a black hole of medicine where no one seemed to agree on anything having to do with a disease that had been dubbed ‘Chronic EBV.’  The problem was that she didn’t have EBV or even the antibodies that would have indicated that she’d been exposed to that virus. Like you, we sought answers, but instead we found confusing and even nonsensical theories about her illness. And thus began the journey for answers.
    • All of us have various milestones in that journey.  Our first and probably one of our most exciting milestones was meeting Doctor Peterson, who at that time was promising that he would do all he could to help Andrea and to help this family.  Our second milestone was really meeting Dr Mikovits. And that was a wonderful and perhaps prophetic meeting.  We were at an international conference, and she had come along as a guest of [The HHV-6 Foundation] when she heard a very important talk about a set of patients who were developing a rare form of cancer.  She took a leap of faith with us to develop a medical research institute when we asked her to come to Reno, and the rest is history.
    • Judy brought with her a scientific passion for discovery of truth, and a curiosity, with well-taught skills from the laboratory of Dr Francis Rosetti, the co-discoverer of the first retrovirus HTLV-1.  Judy has a fiery temperament and a heart of gold.  And when she is not counseling CFS and cancer patients, she teaches students, devises experiments, and travels to major conferences and universities to educate others about the intricacies of the scientific methods used to find infectious and replicating XMRV in the blood of CFS patients.
    • Before I introduce you to Judy, I’d like to ask three things of you. First, please stay involved and advocate for your rights, medical treatment, and adequate funding of research. Your congressmen and senators need to hear from you. Second, please stay informed and educated.  Listen critically to what is said and who is delivering the message. Are they speaking on your behalf? And third, I want you to know that the WPI is going to continue its promised mission. We are not going to stop until we find the answers, but we can’t do it alone, and so we do continue to ask for your support and your help. We appreciate so much all the donations that have come in, all the amazing good wishes that have come our way, and we wanted to thank you today all around the world for letters that have come in to support this effort; that really really helps.
    • So now it is my good pleasure to introduce you to Dr. Judy Mikovits, research director of the WPI.
• Dr Judy Mikovits: 
  • • Well, thank you Annette.  I too would like to thank ProHealth and particularly Kristen Loomis and [The HHV-6 Foundation] for sponsoring this event. What Annette didn’t say is that it was Kristen Loomis who put us together by asking me to attend that meeting in Barcelona, Spain. I never did get outside to see Barcelona, but I saw some amazing scientists and physicians there in the room and in the meetings.  So I’m also honored that you came out on this day, and I understand how difficult it is for patients to get here, and I appreciate all the calls and letters we have gotten around the world since the publication of this paper.  It’s amazing the response that we’ve gotten, and we’re just delighted.  We work for you.
    • [SLIDE 1:  Whittemore Peterson]
      • • The institute is a translational research institute.  You can tell the architects drew this because they’re making a lot of money.  We don’t usually drive Porsche cars [laughter].  But at any rate, this is what the building looks like.  It is three-quarters built, and will be open to serve patients in September this year.  So, keep your eye out for the opening ceremonies and the ground-breaking there.  We’re excited to see patients.
        • Before we could see patients, Annette knew that she would need to start a research program because there were no bio-markers, diagnostics, treatments, or anything.  We started looking with the patients there, with the diagnostic acumen of Dan Peterson.  So, I came up right after that meeting in Spain in 2006 and spent the summer just meeting the patients. I’d never heard of the disease before then, and it was just eye-opening to me to see how sick these patients really were and to try and understand really what the disease was.  My dear husband, when I talked to him about Reno, all he kept saying was “Reno?  That’s not by the ocean” [laughter], but I’m a patient advocate as well in Ventura County with a Cancer Support group with Bible Fellowship Christian Cancer Support Group, and they were kind enough to let me go to Reno because I said “These people are much sicker than you.”  [laughs]
    • [SLIDE 2:  XMRV, a New Pathogenic Retrovirus]  
      • • I’m going to talk to you exclusively about the retrovirus XMRV.  I understand that I’ve left a lot of the detail of the science because, when we see the science, you start to understand the implications of this discovery, in not only this disease but perhaps a number of old diseases where we might find a new understanding. So, this is the Cleveland Clinic rendition of the retrovirus, and of course the electron micrograph that accompanied the publication in Science.  I’m going to give you a little history lesson.  We’re going to walk through the publications.
    • [SLIDE 3:  Identification of a Novel Gammaretrovirus]  
      • • Interestingly, XMRV was identified by Bob Silverman and Joe de Risi in 2006, so just at the time that we were meeting each other and making these fateful introductions, this virus came out, where Bob Silverman, who was an immunologist at the Cleveland Clinic, was looking at prostate cancer patients where there was familial prostate cancer.  That is, it’s hereditary, but that it’s hereditary in a funny way where maybe brothers-in-law or distant relatives would get it and not direct father-son, things like that.  We had a case in my own family where my stepfather died very young of prostate cancer. It’s an aggressive cancer, and when you get prostate cancer very young, it suggests there’s something else going on environmentally.
        • So he looked at a single nucleotide change of a variant in an anti-viral gene known as RNase-L.  This gene, the protein’s job is just to degrade RNA from viruses and protect you, and turn on the interferon response.  But he found a variant in that gene where that single base change, that is in about 13% of the population, makes this enzyme only about 20% as active, so it dysfunctional because it doesn’t work.  So he hypothesized that maybe these men were susceptible to a virus.  And he met Joe De Risi at UCSF, who had a technology which is basically a chip.  It’s like a chip with a bunch of information on it.  The information is just sequences of every known mammalian virus, so 20-30 maybe 70 base pairs of every known virus from a considerable spectrum.
        • He simply took the DNA from these men and applied it to the chip, and the red part you see right here shows that it exactly matched [if you want to match in opposite] the sequences from this particular virus.  So when they took this out, and they sequenced the virus there, they found that there were retroviral sequences in 10% of those tumors, and those with that particular variance, and that those sequences were most closely related to what was xenotropic murine leukemia virus.  It’s a gamma retrovirus, and we’ll talk more about that later.  They just number them alpha, beta, gamma for convenience because the numbers are known.
    • [SLIDE 4:  Xenotropic Murine Leukemia Retrovirus-related Virus]
      • • Xenotropic means it can no longer infect mice.  Xeno means foreign.  So what we know from the xeno family of viruses is that they look like murine leukemia viruses, but they lack a receptor, and we’ll talk about that later, so that they can’t infect mice.  So he named this virus Xenotropic Murine Leukemia-Related Virus because it wasn’t exactly the mouse virus.  Clearly it was something different, suggesting that this might be a new virus.  So, his laboratory did a little more work in the next 2 years.  Again we’re just talking about 2007-2008.  Usually it takes a year just to get a paper published.
        • What he at first identified, they knew that the mouse family of xeno viruses would recognize and bind and actually enter the cell through this receptor.  So it sees the receptor; it’s called XPR-1, and this is a calcium channel type, an ionic receptor.  They don’t know the function of it.  This is called the G protein; it has a particular role in sequencing.  We know there’s a loop right here or so, where the mouse virus has two or three amino acid changes, and that’s why the virus can no longer infect mice.  That’s one of the reasons we know this is not from a mouse.  This receptor is on every cell in the body.  So it doesn’t tell you a whole lot about the infection or what cells would get infected in the disease.
        • The next thing he did was he molecularly cloned this.  He used techniques to write the virus, the entire 8,000 base pairs, and put it in a vector which allowed him to multiply it and make it an infectious virus.  So, he made this infectious virus, but he didn’t actually isolate it.  Using the infectious clone, he then infected various cells and found that the virus integrated.  It inserted itself into DNA preferentially at the start site of genes.  And that’s the part of the gene that turns on and off their expressions, so a lot of the differences you see in patients could be explained by turning on and off the wrong genes when a retrovirus integrates.
    • [SLIDE 5:  Genomic Structure of Gamma Retrovirus]  
      • • Let’s do a little bit of retrovirology 101.  This is the genomic structure of a retrovirus.  Now, retroviruses have an RNA genome.  We have DNA genome.  We have nucleic acids; our genetic information is packaged in DNA.  This virus has a single stranded RNA genome that’s present in two copies in the virus.  So, it first has to be reverse transcribed by the enzyme reverse transcriptase.  So, you have to take the RNA back to DNA and then the integrase gene there, shown here; this is a pol, so all a simple retrovirus will encode is the structural proteins, gag, pol, and the envelope, and then the enzymes.  They don’t have any extra proteins like HIV or HTLV-1 which are complex retroviruses, and they write a bunch of proteins that regulate different parts of your body.
        • The good thing about this virus is that it’s a simple retrovirus.  There’s less that it can do to interact with your cells to have those go wrong.  That’s the first piece of good news.  It’s the first ever simple retrovirus known to infect humans.  We can think a lot about that as scientists and how it might cause disease.
    • [SLIDE 6:  Schematic of Retrovirus Particle]  
      • • Once you make the virus, you go from your genetic information I showed you in the last slide into the envelope protein which has two proteins actually in the surface unit.  This is what binds to that receptor, and then the trans-membrane unit that sets itself into the matrix of the capsid.  This is the capsid protein there, and that’s known as gag, so you’re gonna see capsid and envelope throughout this talk, so you’ll understand that when you have antibodies that develop.  These antibodies are recognizing these areas of proteins, and this is depicted here as that double stranded RNA nucleus, and the polymerase which simply writes the RNA into DNA and then packages it all up and leaves the cell.
    • [SLIDE 7:  Detection of Retroviruses]
      • • Now let’s talk about how you detect retroviruses, because that’s important in thinking about how we found this virus and how we study it in the laboratory.  The viral life-cycle is as I just described.  Once you have the DNA integrated into the chromosome, once it’s integrated to the reverse transcriptase integrated it into the chromosome it’s there and it replicates every time your cell divides and your DNA replicates.  So if your cell isn’t dividing, theoretically it’s just latent.  It’s just there in the DNA, and it’s not making more viruses, it’s not making copies of itself.  It’s not infecting more cells.  This is a good state, if you have a retrovirus, is to just shut down the transcription.
        • As Annette said, when Magic Johnson was found to sero-convert, they found an antibody in his blood, so that’s very shortly after he became infected.  So they were able to give him therapeutics to prevent the virus from making many more copies in his body.  Theoretically the reason why he never got sick is because he is maintained on those anti-retroviral therapies as well as the immunomodulating therapies.  He’s kept that virus down so that he never theoretically will get AIDS.  We’ll talk more about that later.
        • Once the cell starts dividing and you start writing that DNA and transcribing it into all of the proteins we just discussed, the envelope will then package the core of the capsid there, that looks like this, in the RNA genome, that double stranded genome, and it actually uses your cell membrane, cholesterol, and lipids to leave the cell then and look like that artist’s rendition of the viral particle.  So, when you’re looking for retroviruses, and there are only two known, the HTLV-1 family – there’s a one and a two – and HIV, the human immunodeficiency virus.
        • As Annette mentioned, [Dr. Francis William Ruscetti (born 1943)] discovered this virus and reported it in 1980.  At that time, there was no PCR, so he couldn’t look for an infected cell by a sensitive method.  He looked for that enzyme reverse transcriptase because reverse transcriptase is only in retroviruses and not in human cells, so it’s easy to look for the activity of that protein that would then transcribe and make the virus.  And sometime maybe if you’ve learned the history; it’s amazing the small small signals they found in the early days to describe the virus.
        • But then you’re going to do what we call a Western Blot, which is to run out the proteins of a cell on a gel electrophoresis and just blot it and look for antibodies – and we’ll show you those later – for the viral proteins and test for specific antibodies to the envelope and the gag proteins and just look for the presence of virus in infected cells. The first thing you do clinically is you look for serology – that test that shows you that your system is making an antibody to that virus.  That was the test that Magic Johnson got.  You have a virus in your body and your immune system’s job, to distinguish self from foreign.  So we know this is foreign because you have made an antibody to it and then finally, it’s rarely done clinically, to identify HIV or HTLV-1, is to isolate the virus and actually purify it in cell culture.
    • [SLIDE 8:  Detection of an Infectious Retrovirus…]
      • • So that leads us up to the next paper.  After the first bit of work that Bob did in describing this virus, there wasn’t a lot of excitement about it in the scientific community, because they didn’t know that it was an infectious virus.  It was just sequences in prostate tissue tumors, and it wasn’t meaningful to the scientific community because we all have sequences of viruses in our body as we all know, maybe as much as 15% of our genome is made up of viruses that are silenced by our immune system so that they can’t be expressed.
        • The work that we did then generated a lot of excitement.  What we did was we detected this infectious retrovirus and showed that it was infectious in the blood cells of patients with Chronic Fatigue Syndrome.  We’re going to walk through exactly how we did this to show the virus. At first we did PCR because at the time this paper was done, the only thing that was known was Bob Silverman’s specific PCR technique.  We had not validated or identified any antibodies.  It was not known that it was a pol-virus, an infectious virus.  So that’s what this work was, it was serendipitous really, that we happened to have the patients who had this virus because, if we did not have a well-identified cohort of CFS patients, and we were just looking at the general population, retroviruses aren’t highly expressed in the general population.  HTLV-1 is 0.2% in the US population, and we’ll talk a little bit more about what that means too.  Retroviruses are not ubiquitous.  It’s not like EBV and CMV, where everybody has them.
        • We had these well-characterized patients who had been sick for many years.  I think it was a large part of why we’re able to isolate this virus.
    • [SLIDE 9:  CFS Study Cohort Reported in Science]  
      • • We’ll start at the beginning, and that’s the cohort, who they are.  When it came to the Institute, what we talked about, was really important, was having a repository of samples from all of the patients so we could look at the RNA for their gene expression, at the DNA for maybe what was different about the genetics of some of you that might make you sick. Then we look at the plasma for proteins to see if we could identify immune modulators called cytokines that tell your immune system and tell your brain how to function.  So we made these samples across RNA, DNA, protein and plasma, and then a culturable cell, so we kept some frozen such that we could grow them up and make more of them, any time we wanted, of your peripheral blood mononuclear cells; that’s your white blood cells.
        • We used patients who came literally from around the world, and this was actually not correct in the Science paper because I didn’t know there were international people in the repository at the time.  When they come to Incline Village, it’s assumed that they are from Nevada, and when we decoded this over the Christmas holidays we found 12 or 15 states, the UK, Ireland, Germany and Australia as well.  So we had both international and people literally from all over this country, not necessarily Reno, Nevada, where the associated outbreak that we know occurred there in the early ’80s.
        • The conclusion… all you had to do to be a sample in our repository was have a CDC diagnosis of Fukuda criteria or the Canadian definition diagnosis which is more stringent for various immune defects and inflammatory defects.  Regardless of severity, the samples in the repository are from people aged 19-75.  We don’t have any whole bodies yet of people; though people do offer to donate whole bodies, however I don’t think we need them at this point [laughs].
        • The study characteristic, like the disease, was 67% women, reflecting the gender bias in incidence of CFS.  The mean age was 55, but some of these people had been sick since they were children or early 20s or early 30s, so they had a long haul with this illness. The 218 control samples were de-identified samples, so we don’t know who these people are.  They came from two places; they came from a medical practice in Reno, they came from a doctor who identified these people as healthy, and these were collected from people before I came to the University in 2007, and they were looking at the immune systems of healthy people to identify some of the functions, so we were able to use those samples under IRB approval.  There is also a paternity diagnostic company in Reno, where they get samples from all over the world from mom and dad, so we tested from those 100 or so samples too, so we were at least able to zip code match.  We have regional areas for the geographic location, so it was matched for location.
        • This is a PCR gel; I simply run them out for electrophoresis, and that gives you a different size so you can look at the exact size of the fragment of DNA that you’re looking at.  Again, this was done by Bob Silverman who is our collaborator in the study.  Today is actually the first anniversary of January 22nd when I called … we saw some of these data right after the Christmas holidays, and we had promised Bob for a long time that we would look at this because RNA cell is a major defect in our patients, whether it’s underactive or overactive.  Something is wrong with the RNase-L pathway in CFS patients, according to decades of research.
        • We promised Bob that we would simply look, although we had done micro-ray technology, and we had not found the virus there.  We had his specific primer pairs so we could go in and look for that gag structural and that envelope gene, so that we could see viral sequences in the cells that could make viral proteins theoretically.  What we found was that 67% of the patients we looked at, we could find sequences in both the gag and the envelope gene or just the gag depending on the virus life cycle at the time.  This was astounding because we only found the sequence in 3-4% of the healthy control population.  It’s also interesting; I said 68 out of 101 patients.
    • [SLIDE 10:  Presence of XMRV Sequences in Human DNA]  
      • • On some of these patients, we looked three and four times for the DNA in the unstimulated cells.  So this is just that pellet that I made when I sorted all the various samples.  I just held one as white cells so that I could make DNA later or RNA later, depending on the technique I wanted to use downstream.  So, it’s important that this was in 68 out of 101 samples.  It was 68 out of 101 patients, and it clearly says that in the paper.  At any given time, depending on the viral life-cycle, we might not find this virus in the unstimulated group (inaudible).  And I give you the example of that is: follow this patient 1118 throughout the talk, and you’ll see that this patient, if you only use sequences, would have been called ‘negative.’  So, we were concerned because PCR is a technique that is fraught with contamination.  If you’re looking for a needle in a haystack, just a few sequences in a million bases, you might make an error in your enzyme, and it might put the wrong base in there.
    • [SLIDE 11:  Comparison of Nucleotide Sequences from XMRV…]  
      • • Jaydip Das Gupta in Bob Silverman’s lab cloned and sequenced three of these patients – and that’s shown here – and what it’s intended to show is: If you compare the isolates that they had from the 3 prostate cancer cases, where they had actually cloned these, you can see, if you compare it to the reference strain, known as VP62, that’s the reference strain of what this virus looks like, the CFS samples here were clearly different, but they were highly similar – 99.7% – there were maybe 8 bases different across the entire 8,000 base pairs. This virus isn’t like HIV theoretically.  It’s not changing.  We don’t find quasi-species in patients when there are lots of different viruses, because HIV mutates so much.  Therapeutically, that’s something that we can take advantage of and suggests that it might be easier to develop therapies because the virus is going to be largely the same.
    • [SLIDE 12:  XMRV Isolates from Prostate Cancer and CFS…]
      • • Rachel Bagni, my former student at the National Cancer Institute – I asked her if she could construct what is called a phylogenetic tree of this virus so we could understand where it came from (hopefully).  And so that’s shown on the next slide.  And what a phylogenetic tree is – is you take all of the sequences of all the Murine Leukemia viruses – they’re called Ecotropic viruses – all the families of virus that they’ve ever identified, Mason-Pfizer Monkey virus, all the sequences, and you put them into the computer, and then you put into the computer at the same time the sequences of our 6 isolates – the 3 prostate cancer and the 3 CFS isolates that we had at that time.  And you do what’s called ‘blasting’.  You ask the computer to find similarities.  And when it doesn’t find similarities, you get what’s called a new branch on the tree.  So, clearly, these diverge here, and we don’t know when that is in time, but these data suggest that the prostate cancer – that XMRV both in prostate cancer and in CFS – form a new distinct branch, that it’s a new human retrovirus.  It doesn’t have any of the sequences of mouse in it.  And when we blasted it, also we did the same thing against the human genome – because I told you, we have a lot of endogenous viruses that don’t actually come out of our bodies as infectious particles – we blasted it against the human genome and found that it did not match any sequence in the human genome.  So, it’s clearly a foreign, exogenous virus that can now, theoretically, be infectious.  And that’s what we’ll show in the next slide.
        • Here are our sequences.  And you can see, they are clearly not contaminants.  We didn’t have this – we weren’t working with this in the lab, actually, at the time.  But we didn’t have this, and maybe spread it through the sample in any way.  It was there – clearly different isolates.  We now have more than 170 isolates, because we isolate from every single patient in all of our studies.  And we’re actively looking for funds and going to sequence those viruses because it might give us clues as to some of the differences in what we see, maybe something, you know the various symptoms, because CFS is quite a heterogeneous disease.
    • [SLIDE 13:  XMRV: A New Human Retrovirus]
      • • At any rate, we next went to – I’ll summarize that – in summary, what is XMRV then?  These data suggest, at this point in time, we have sequences related to XMRV that were not found in any mouse strain.  So, it’s a new human retrovirus.  The origin of XMRV remains unknown.  We don’t know how it got into the human species.  We don’t know how long it’s been – 40 years is the guess of John Coffin, who is a mouse retrovirologist working on these families of viruses for more than 40 or so years.  And that XMRV is not a mouse virus – clearly from these data.  So it’s a new human retrovirus.
    • [SLIDE 14:  Detection of XMRV Protein Expressed in Activated…]
      • • We next asked: Could we find those proteins I mentioned?  We took advantage of (inaudible).  Sandy Ruscetti, Frank Ruscetti’s wife, had been in retrovirology as long as he had, but because they didn’t want to work on the same thing, men usually get the credit for what women do.  So Sandy worked on mouse viruses, and Frank worked on human viruses, and I don’t think they actually ever published together.  But we were thinking about it and saying: None of the reagents that were out in the world, so far nobody had found viral proteins from XMRV, even though it had been discovered two or so years earlier.  In January we started looking.
        • Sandy had saved a box of antibodies – this is really a tribute to the value of your tax dollars going to basic research – because they created this mouse retrovirology program and put a lot of money into trying to understand – if you can understand how viruses cause cancer in mice, you might understand how it causes cancer in humans.  And this was in the late ‘70s and early ‘80s.  And somewhere in the early 2000s, they were going to throw out all of these reagents that they developed and Sandy said, “No, I’ll keep them in my freezer.”  Frank always says that the reason they’re still married is because Sandy never throws out anything.
        • At any rate, she gave us these viruses, I mean these antibodies, and we screened our samples there for protein in our samples.  So, we looked at the activated peripheral blood mononuclear cells.  And what we do is we stimulate these to divide, and add T-cell growth factor, or now known as IL2, which was actually the discovery that Frank made that preceded the identification of the first human retroviruses.  Retroviruses grow and divide in cells, so you have to divide the cells in order to get the virus to replicate to levels that you can see with the technology of the time.  And that’s important in this study too.
        • What we’ve got here is we looked at a number of her antibodies – these are all family members of the virus – this particular antibody which you’ll hear a lot about is a spleen focus forming virus.  It’s a mouse virus that causes various diseases including a neurological disease and erythroleukemia – red blood cell leukemia.  Its envelope is both a neurotoxin and an oncogene.  It causes cancer and causes toxicity.  This virus itself – she had this antibody that was highly specific.  It recognizes all known polytropic and xenotropic viruses.  We hypothesized that it would recognize this virus and clearly high levels in some patient’s cells, but not in others.  Interestingly enough, if you look, and use a panel of antibodies, this is a gag antibody to a gag protein I showed you there, that structural gene and this virus, this antibody is a polyphone virus that recognizes the entire MULV.  And you can see when you use a panel of antibodies to the viruses, essentially everyone, 68% now of 50 people we tried just one time, you could see their proliferating blood cells.  You can see evidence of viral proteins.
    • [SLIDE 15:  Absence of XMRV Protein Expression…]
      • • We next asked if we could see this in normal cells, because of course you want to make sure that it’s not in normal people.  And you can see clearly here in the 24 normal donors (now up to 60 or 70 that Frank’s done) at the NIH clinical center where they have a good donor program – they’re all negative.  So, these proteins, these viral proteins are expressed specifically in the CFS patients and not in normal donors.
    • [SLIDE 16:  Transmission of XMRV from CFS Patients…]
      • • We next asked if we could transmit that.  Is there any evidence that it’s an infectious virus?  So the first thing we did was we took plasma – so that’s the plasma, the liquid off the white blood cells there – and we took their plasma and [this becomes essentially the key to the whole study] we co-cultured it.  We simply put it in a flask with the cells known as LNCaP and that abbreviation comes from lymph node-cancer-prostate.  This came from a lymph node of a 62 year old man who had metastatic advanced prostate cancer.  And these cells grew by themselves in the laboratory so that you could use them as a tool for studying prostate cancer.  And, in one of my lives, I developed prostate cancer drugs, because, when my stepfather got ill, I became interested in prostate cancer and had been working on this.  So, I knew LNCaP was also deficient in RNase L, and the type one interferon pathway.  It had no interferon response.  So, we always look for biological multiplication of the virus instead of the multiplication you would use with PCR.  So, actually replicate the virus or multiply the virus in cells.  You have to find a cell that will grow a lot of virus so that you can study it.  So we took that plasma from all of these patients.  You see high levels – now 84% of the plasmas contain infectious virus that we could not see.  I sent all of these plasmas to Bob Silverman and he said, “Sorry Judy, I don’t see the RNA of the virus” there when he looked for the two copies of RNA in the particles which suggested there were very few copies of actual particles of virus in these cells.  But again, we could transmit it.
    • [SLIDE 17:  Transmission Electron Micrograph…]
      • • And the next question we asked is: Is this a whole virus?  Is this an infectious virus?  Kunio Nagashima, my friend at the NCI who is an expert in Electron Microscopy, did this electron micrograph for me, and what you can see here is the budding of a virus from the cell.  It shows you again that it’s not a contamination, it’s actually a transmission, because you’ve got a budding particle.  And that particle is called a C-type retrovirus, because in the old days, when we used the word, they called them ‘C’ but they changed the name to gamma, but we’re old-fashioned, so we keep the ‘C’ type.
        • And what you can see here, characteristic of a gammaretrovirus, you can see this budding – remember I showed you it takes the cholesterol and buds itself out of the cell to form the outer membrane.  And here’s that capsid that encloses where the viral RNA is, to protect it.  So you can see both immature particles and many mature particles in those LNCaP that have just been exposed to patients’ plasma, showing there is an infectious virus there.  So the next thing, so we were pretty happy with this and we sent it off to Science in early May of last year, and they came back to us and they said, “We’re 95% convinced, but show us an immune response.  If this really is an infectious, non-self virus, not an endogenous virus, your body will make an immune response.”
    • [SLIDE 18:  Antibodies in CFS Patients’ Plasma…]
      • • Again we went to Sandy Ruscetti, and this part was funny too because we were struggling to do this, because you don’t want a whole virus infected cell, you need to have just a part of the virus in order to get the noise out of there.  And what Sandy had developed when she was studying the spleen focus-forming virus was this antibody again to the envelope protein.  And she expressed it on the cell lines – used two cell lines.  This is a mouse b-cell line that expresses the erythropoeitin receptor (it’s just for red blood cells), and when she co-expressed the envelope, you see high levels of the envelope on the surface of these cells.  So we took these cells and put them in what’s called a flow cytometer, where a laser will see the fluorescently tagged antibody on the surface of the cell and count the infected cell as it runs through the instrument, the channel and single cell.  So you can see that the cell line went out the envelope protein being expressed.  You see the white and the black are superimposed showing that there’s nothing reacting specifically with that.  If you then take that antibody I showed you, to the envelope, its called 7C10, and expose the cells to it, they all light up, virtually 100% of these cells have the antibodies that are recognizing the cells with the envelope protein.  If we then take a patient sample and do exactly the same thing, you see there’s an antibody, this is for patient number 1104, that’s one of the sequences we have, and there it is, there’s the immune response in the plasma, showing now we have an infectious virus with particles that can exogenously infect and is non-self.
    • [SLIDE 19:  XMRV is Present in Malignant Prostatic Epithelium…]
      • • The next step in what happened in the literature is work in prostate cancer again.  So this comes from the lab of Ila Singh, who’s an MD-PhD at Utah, and she was looking at XMRV in malignant prostate cancer tissue in the tumor cells.  One of the other reasons why the oncologists in the cancer community weren’t excited about Bob’s discovery of XMRV sequences was because, when they looked at those, they only found them in the infiltrating stromal cells – the microenvironment.  But those of us who think a little deeper than most oncologists about cancer know that 50% of all tumors are actually your immune system, your white blood cells, going in to try and clear the cancer because that’s their job is to recognize tumor cells.  So we weren’t concerned.  We were excited that it was, and it made sense to us that it wasn’t the tumor cell itself harboring the virus, but the immune cells that were inside the tumor.
    • [SLIDE 20:  Fluids from XMRV Positive Prostate Cancers…]
      • • But Ila showed that XMRV WAS present in the malignant tumor cells and that it was associated with that high grade tumor, that tumor that my stepfather died of, that you get younger, and they get really sick really fast.  And what was different in the advance in her study is she developed an antibody specifically to XMRV, to the whole virus, another polychromal antibody.  And she showed that she could recognize with that antibody, in what’s called Immunohistochemistry.  When you send a biopsy to the lab, they look at it, at a tissue block.  So she did that, and she showed that 23% of the prostate cancer tissues she looked at had a protein to XMRV, a lot like our study, but she saw a lot less DNA sequences than she saw proteins.  This paper came out about a month before our paper, but we knew about it from about mid summer when we first met.
        • In her study, the limitation in her study, was that again that there is no evidence of the infectious virus that I just showed you.  So we had evidence of infectious virus in CFS.  Can we see evidence of infectious virus in prostate cancer?
    • [SLIDE 21:  Transmission of XMRV in LNCaP]
      • • Frank did this, this is again that antibody, looking for the antibody in the patients.  And here he used, this is called a prostatic secretion, so they’re just looking at the prostatic secretion and when they had a person who had sequences of the virus, positive in the prostatic secretion, you can see there that there are antibodies in that patient, so that patient is infected.  In an XMRV PCR negative patient, we don’t see antibodies, so that person is unlikely to be infected with XMRV.  And again in the plasma of this integration here, so that now they have actually found in this patient exactly where the virus integrated into the cell, and that patient has a significant amount of antibody.  In prostate cancer, no one had ever transmitted virus and shown that it was infectious that way.  I show you the exact same study, where we took the plasma from the prostatic secretions there and found high levels of the virus when we put it on LNCaP, showing now in both prostate cancer and CFS, that XMRV is an infectious virus.  And in a significant portion now they are finding in prostate cancer patients.
    • [SLIDE 22:  PCR is not as Sensitive a Detection…]
      • • Why bring that up today, is because if we look and we do a summary table of the technologies that I showed you that we used to find the virus, what you see is that patients here in red are clearly infected when you look at plasma antibody responses, and you look for transmissions through infectious particles in the plasma, you can see the red patients both in the prostate cancer and in the WPI patients.  These patients were PCR negative, I bring back to you 1118, but we found plasma transmission of that virus that I didn’t point out, pardon me when we passed that slide.  But ALL of these samples were negative when you did the most sensitive PCR that Bob and everyone developed in unstimulated cells.  So those white blood cells, fresh out of the body, not dividing, very low copy numbers of this virus, but clearly these individuals are infected.
    • [SLIDE 23:  What is the Incidence of XMRV in Prostate Cancer and CFS]
      • • Going back to the literature now, two studies have come out since then, and one was in October, right around the time our paper came out.  And this was from a German group led by Norbert Bannert, and he found a lack of evidence for the virus in over 580 prostate tumor tissues, when he used the sensitive nested gag-PCR techniques that me and Bob and everyone is using right now.  And he had developed his own ELISA which is looking for an antibody in the sera – it’s a similar test to what I showed you for looking for antibodies to that.  And he couldn’t see any of the evidence of the virus in those sera, and so he concluded, they concluded that XMRV was not in prostate cancer.  And then earlier this year, a similar study came out by a group in England that showed a failure to detect XMRV in CFS.  And they looked at 186 DNA samples, and they did nested gag-PCR and they found nothing.
    • [SLIDE 24:  What is the Reason for Discrepancy]
      • • What could be the reasons for the discrepancies in these studies and what we’ve shown you in the studies of Ila Singh?  First of all, the prevalence of XMRV, that’s the distribution around the world, is unknown.  The studies that we’ve shown you today is all we know about XMRV prevalence – that it’s in the US and in several hundred people including those with both prostate cancer and CFS.  But I remind you that retroviruses are not ubiquitous, they’re not everywhere.  The sensitivity of the assays in these studies was not the same because both of these studies didn’t rely on (inaudible), they relied on PCR, they didn’t look for infectious virus.  Of course the Bannert group didn’t know our study because the papers were under consideration at the same time.  And then also that XMRV has an extremely low copy number that I showed you, that even if it is there, you could miss it by these sensitive techniques.  And mostly importantly, and something that didn’t occur to me until I saw all of this data, was that we don’t know anything about the viral reservoir of XMRV.  I assumed it’s lymphocytes because that’s what I know about HIV and HTLV-1.  But what if the plasma virus was coming out of the tissues, and then the cells that were actually in the peripheral blood were not the main reservoir of the virus?  What if there is another tissue reservoir?  We don’t know what that is, so these are all possible explanations for why we saw it, and we see a lot of it as you see in the plasma of these people, not a lot by copy number, but certainly there is infectious virus there.  So that’s what we’re thinking.
    • [SLIDE 25:  Distribution of HTLV-1]
      • • If you look at data that supports these arguments, what you will see is the distribution here of HTLV-1.  Now HTLV-1 infected people are 10-20 million across the world, and I bring up this one point that HTLV-1 causes a neurological disease known as HTLV-1 Associated Myelopathy.  They have trouble walking and balance and almost like a paralysis looking disease.  And that occurs only in about 20% of the infected individuals.  And then of course HTLV-1 was shown to be causative, satisfied Koch’s postulates as we know them for viruses – for an adult T-cell leukemia, and this is a very aggressive leukemia, and the mechanisms for how it causes that are still largely unknown.  But at any rate, 10-20 million people are infected, but you see very few – only sporadic cases occur in the US or Europe, and the US incidence is only about 0.2%.  They don’t even test for it in the blood supply because it’s just simply not a problem in America, its endemic in the regions that are shown here today.
    • [SLIDE 26:  Transmission of XMRV from Activated PBMCs…]
      • • And the second argument that supports maybe what’s different between these studies is the transmission from the active PBMCs.  So if I take the white blood cells, some of which where I can’t see virus and just put them on LNCaP, I can transmit the virus to this indicator cell-line that has shown you because its defective in RNaseL (theoretically because its defective in those, but we learn more about it later), will amplify and replicate high levels of the virus.  There are scientific reasons why there are differences between these studies, but I don’t think there is any doubt that XMRV is a new human retrovirus, and since both HIV and HTLV-1 are associated with neurological diseases and cancer, and now we have associated them with a neurological disease and cancer, that this is a real human pathogen.
    • [SLIDE 27:  Recent Publications – Clues to Pathogenesis]
      • • Recent publications after those publications (I’m just walking through the literature off the last few years) might give us a clue to the pathogenesis – how XMRV might cause disease.  So this paper by Steve Goff’s lab shows that XMRV establishes in an efficient infection, and spreading infection, that’s enhanced by transcriptional activity in prostate cancer cells.  And what that means is, I told you the receptor is on every cell of the body, but clearly every cell doesn’t have the machinery necessary to replicate the virus to high levels.  In fact we see that the peripheral blood mononuclear cells really don’t, and that’s why we don’t know where the tissue reservoir is.  So he simply infected a lot of different cell-lines, and he found that the expression was very very low level, except in essentially one cell-line, and that’s LNCaP.  So we got very very lucky in that this was the only cell-line I thought about as an indicator cell-line.  We could have screened the hundreds of cell lines I know of that we do regularly when we’re looking for viruses because if you can’t grow it you can’t study it.
    • [SLIDE 28:  Hormones and Inflammation Increase Replication of XMRV]
      • • LNCaP turned out to be really serendipitous, and I think the key technical advance to being able to make that discovery, it’s just clearly luck.  He showed that LNCaP responds to androgens, I told you it lacks interferon and RNA cell anti-viral responses, and I’ll show you what’s called the promoter, the response elements, that might give us a clue as to the pathogenesis.  And then Bob Silverman’s lab showed the same thing.  He showed that androgens stimulate transcription which is the replication and division of the virus.  Here’s a clue to the disease because we know the only two diseases so far that are associated with this retrovirus are prostate cancer (a hormone responsive disease) and CFS (one that’s thought to occur primarily in women).
        • Interestingly that I didn’t say that I knew LNCaP is androgen responsive.  So you can make it do a lot of good things and that’s why we use it in drug development for prostate cancer.  Let’s look at that organization of the gag, pol, and envelope of this simple retrovirus.  This U3 region is highlighted because this is sort of the on/off switch of the virus.  This turns it on to make more of the particle in your genome, so this signals your cellular machinery to start making more virus, and what Steve Goth’s lab showed (and he graciously gave me these slides about mid summer) was that there’s only three responsive elements that turn on this virus that he can find so far.
        • Two are called glucocorticoid response elements and they’re shown here.  When a protein actually recognizes that exact sequence and sits down, it tells the virus to turn on replication.  And so interestingly enough, what turns on the virus?  Hormones.  Progesterone, androgen receptor, and testosterone, and we don’t know all the other hormones.  There are a lot of estrogens and estrogen-like compounds, even in our environment these days, which might tell us maybe there’s an estrogen compound that’s not a naturally-occurring, estrogens in a plastic, in the environment, that is actually turning on the virus.
        • We don’t know all of the things that turn it on at this point.  And the other thing that turns it on is cortisol.  What is cortisol?  It’s the stress hormone, and so right there you’re turning on the replication, so it’s an on/off switch for the virus with the stress response.  When you’re told that you respond poorly to stress, there might be a reason for that if you’re replicating a retrovirus!  (laughs).  Sorry I shouldn’t laugh.
    • [SLIDE 29:  Clinical Research Findings]
      • • Then we went back into thinking about this virus, we thought about the clinical research findings that had occurred throughout laboratories around the world throughout the years.  What it mentioned in part was we know that CFS is a multi system disorder (and in Spanish I say sequelae), but there’s lots of inflammation going on, you have allergies, multiple chemical sensitivities, there’s a lot of inflammation, and increased numbers of activated T cells, and the production of these inflammatory molecules I mentioned known as cytokines and chemokines.  Also a key dysfunction in the immune system of CFS patients is this low natural killer cell activity and sometimes numbers.
        • The natural killer cell has two jobs in the body: kill tumor cells and kill virus infected cells.  In CFS, it’s long been recognized (I think first identified by Nancy Klimas and her colleagues more than 20 years ago) that natural killer cells in CFS patients don’t function normally, although the dysfunction is not known, but that again gives us a clue to the pathogenesis.  So this suggested to us that this chronic infection with a retrovirus (retroviruses are associated with immune deficiencies) might lead to the creation of actual immune deficiency that has patients susceptible to opportunistic infections and more likely to develop cancer.
    • [SLIDE 30:  Hypothesis of XMRV Pathogenesis]
      • • I’ve schematically drawn our hypothesis on the next slide, and I basically just lifted the graph of what happens in HIV and changed it to what we know happens, and changed it to all the data that we have so far.  In HIV what happens is that there’s an early infection, the green line is actually the plasma viral load, and it goes up in a spike.  This might be a flu-like syndrome, or it might be nothing at all.  You might never know that you were actively infected at this point and get sick.  But then you have multiple other infections, stress hormones, advanced inflammatory responses that cause these various spikes of the virus throughout a time course which we don’t know.
        • I’ve heard the incubation period of HIV virus is 21 days.  We don’t know anything about the incubation of this virus, we’ve just discovered it!  At any rate, all these events operate to set the viral load higher, because every time you divide a cell, that your white blood cells, the cells in your immune system, and actually our paper shows it’s the TB and NK cells are infected.  Those cells are getting infected, more and more and more of them, and some of them are long live memory cells that you need.  Or they’re going to the tissues then, and they’re infected, and they’re spreading the virus to other cells, and we don’t know where that tissue reservoir is, and as I said the receptor theoretically is on every cell.
        • Not every cell can replicate the virus, but virus can get into every cell.  So it’s infecting more and more NK cells, as the load keeps coming up, and at this point, something happens to your NK cells.  This envelope antigen comes to very high levels, like we see in our patients’ plasma and white blood cells, and we know that in animal models or in animal viruses of this family is actually a noctogene and a neurotoxin.  We hypothesize that the envelope alone is creating some of the neurological sequelae, and that they’re different from the virus replicate.  So it can be sort of the envelopes around a lot more, I showed you the defective particles with less infectious virus and more defective virus, but those proteins can affect your body.
        • We know you’re making antibodies, but some of the sicker patients don’t make antibodies, and CFS patients are known to have problems with antibody production for whatever reason.  We’re not saying that’s direct to the virus, but you know it’s not a great leap of faith because that’s what we saw in the early 1980s with AIDS patients.  We had no idea how long those men had the virus.
        • All of a sudden, they were getting Pneumocystis and Kaposi’s sarcoma (a form of cancer that only occurs in older men in Italy), and that’s because, as you age, your immune system loses effectiveness too.  All of a sudden we’re seeing a virus that is not endemic in the United States.  Well actually from these patients, they actually identified HHV8 (Human Herpes Virus 8) which actually is causative for Kaposi’s Sarcoma, and that virus, I led a drug development program about a decade ago just before I came to California, and we were going to make drugs to target AIDS-associated malignancies, and we found as soon as we got the highly active anti-retroviral therapy and got rid of the HIV and silenced that, the Kaposi’s Sarcoma went away, as did the HHV8.  They cut the budget for that drug program and rightfully so, because there’s no need to develop these drugs because they learned that, at that point, all you have to do is control the retrovirus, get the immune system back to functioning, and also the good news is most of those men, their immune systems are functioning well.  You can get a lot of them back to at least a level of health, even though they have to stay on various drugs the rest of their lives.  At least they could cure the immune deficiency.
    • [SLIDE 31:  XMRV – A Human Gammaretrovirus of Unknown…]
      • • In summary, then, of the science part of the talk:  XMRV is the first simple human infectious retrovirus.  It’s a gamma retrovirus; it’s not complex, so it’s the first one known in this family, and we know nothing about the pathogenic potential, other than the two diseases that we’ve seen it in.  We know that human retroviruses are not ubiquitous.  I’ve shown you the distribution can be quite low in various places in the world.  We don’t know how it spreads across continents.
        • They’re not benign, meaning they cause disease.  All three known human retroviruses are associated with neurological diseases and cancer.  And importantly, they are not airborne, retroviruses are not contagious, you don’t get them in the air.  We know that, for instance, with AIDS patients, that it’s not a problem to kiss AIDS patients and hug AIDS patients, and so that knowledge is there for this virus as well.  So there’s 3 known now, the complex and now the simple, and I’ve mentioned that a number of times. Interestingly, and something we should think about, in light of the replication studies and the other studies as we’re going on, I say HIV and HTLV, but I’ve been saying one, but there are variants of HIV.  There’s HIV2 that is less pathogenic, there’s HTLV-2 that is less pathogenic, in fact hardly pathogenic at all.  And these are clearly different and have different pathogenic profiles, and just a short extension of that suggests that there could be variants of XMRV.  There could be subtly different sequences of viruses out there that are associated now with different phenotypes, so the way the disease looks, and different cancers or different neurological diseases.
        • I know that the scientific community is actively looking for variants, so another good news about these studies is that there are a lot of excited retrovirologists and immunologists who started as soon as they learned this in July, to the put the world resources and the best minds on this virus associated with CFS, and that’s probably the first time that’s happened in the world, so they’re excited about that.
    • [SLIDE 32:  Personal Reasons to be Tested]
      • • A lot of the questions that I got, and I wrote this talk around the question that I got, had to do with reasons to be tested.  You know we don’t have the best diagnostic test yet because we still haven’t validated that serology test.  That serology test is done in a laboratory; it’s very cumbersome, we need to validate it clinically in order to look for antibodies in the population against this virus, and that is the number one test when you go look for HTLV.  But that said, there are opportunities to get tested, and you might have your own reasons to get tested.  Now generally a physician won’t test because there are no treatment options.  There are no known anti retrovirals currently that are known to be good for XMRV, so why go get a test for it if you can’t treat it?
        • But it can give you additional validation that your illness is an organic illness, and that can have a huge psychological boost because you can begin then to think about immune support and things you might do and changes in your lifestyle where you may be able to support your immune system in the meantime while we develop drugs.  And importantly, you want to protect your personal family and public health.  We need to know where this virus is.  And it does help, physicians then start to see, physicians like Dr Peterson, will know how that might relate to your other infections, your other immune issues, if you have cytokine profiles in some of the tests he does.  It might help him or some of the other physicians with your therapy to know that this is a player in the game now.
        • And again it underscores the more people that are infected, that 3.75% is 10 million Americans, so that I didn’t have to say anything.  The drug companies called me the next day and said “Gee we’d like to help!” and so we’re actively working with them, and they are helping because there’s another piece of good news which is that there are drugs that were on the shelf that were developed all the way through phase 2 clinical trials.  They were shown to be safe in people, but they just didn’t work as well against HIV as the drugs that were out there.  Why spend a lot of money developing them?  There are real targets that you can go after that can serve regions between these viruses right now and maybe come up within the next year with a drug and a clinical trial for that drug that would go a long way toward treatment.
    • [SLIDE 33:  Preventing the Spread of XMRV]
      • • Right now we recommend, to prevent the spread of XMRV, if you have CFS and you wanted to be prudent, even if you didn’t get tested say “Okay I might be infected.” What would we recommend? The HIV precautions, because it’s a retrovirus we know it’s spread, we found it in blood, in the body fluid secretion, and prostatic secretions. You just want to assume that these precautions that are very stringent, and have prevented the spread of HIV in some countries, that if you don’t donate blood or sperm (this virus can infect sperm cells), so if you have CFS or maybe a history of aggressive prostate cancer in your family, you might think about not being a blood donor.
        • Follow the HIV precautions. Don’t share toothbrushes, because you can have bleeding gums or razors. Use safe sexual techniques, and do not breastfeed. It’s ‘don’t breastfeed after six weeks when the maternal antibodies go away.’ When they did that in Japan where ATL (that aggressive leukemia) was rising in the late 70’s and early 80’s, all they did was say “Okay, no breast feeding!” and 40% reduction of ATL rates in Japan. So prevent the spread of this virus, and you can reduce the disease and protect your family.
        • Part 2: Transcript with slide citations of the Jan 22 Santa Barbara XMRV Seminar by Whittemore Peterson Research Director Dr. Judy Mikovits – compiled and edited by long-time ProHealth subscriber and contributor Sandy Miarecki. (To return to Part 1, click here. To download the slides, click here.)
    • [SLIDE 34:  WPI Research Priorities]
      • • What are our research priorities? At the WPI, we’re actively working with the federal government to develop that next generation of tests. We expect that serological assay (Rachel will get mad at me) within a month. She told me yesterday that the data were looking really good. And we want to investigate the prevalence of XMRV. The federal government – the National Heart, Lung, and Blood institute – actually called as soon as the paper came out, and we set up a Blood Working Group to investigate what is the true prevalence. Prevalence means the presence, the distribution, not necessarily the disease; and we use incidence with disease, and prevalence of XMRV in the blood supply. Our numbers were small – they were only 200 or 300, that’s 4%.  But 4% is still 10 million Americans, so you want to look at that, and they actively are.  And they’re working on that second generation test as well.
        • We want to understand those tissue reservoirs, and clearly it may not be the PBMCs (peripheral blood mononuclear cells). Is it the lymph nodes?  Is it bone marrow?  It’s possible (I don’t expect it), but it could be the brain.  We don’t know at this point. We are actively working, as I said, with drug companies to develop anti-retrovirals and immune based therapies.
        • We want to understand how it’s transmitted.  We’ve got a family study going on in the research plan; it’s just getting IRB (Institutional Review Board) approval and ready to start.  Hopefully we can get families who have any number of diseases across the spectrum, fibromyalgia, other neuro-immune diseases, maybe a higher incidence of cancer, but we need healthy people as well, so we’ll take essentially anybody into that protocol.  And, as I said, that protocol will help us investigate the incidence of XMRV in other neuro-immune diseases.
    • [SLIDE 35:  Questions]
      • • Important questions that the field is working hard to answer (and we are as well, but we won’t be able to do all this):
          • • Is XMRV a causal factor in CFS, and possibly some aggressive prostate cancer? And we’ll talk a little bit about how you think about a causal factor. One way to do that is, we have several patients who came to Dr. Peterson, and they said “I was fine until I got into a car accident. I got a blood transfusion in the hospital and I got CFS” or “I had a surgery and had a blood transfusion.”  So if you can identify a blood transfusion exchange of an acute infection that causes the disease, and the virus wasn’t there before in the human, and it’s there afterwards, and it’s in the donor, then of course you’ve got causality, and that is one way that causality was shown in HIV as causing AIDS.  I think we all know the tennis player Arthur Ashe, and that is how he got HIV/AIDS and subsequently died.
            • How does XMRV enter the human population? Is it a zoonotic [from animals] transmission? We know it’s not a mouse, at least not any of the mice we know. It could be a field rodent of some kind, but we’ve never found the virus in another animal. This is the first animal that is the “Xeno,” and that is man. So, how does it enter the population, and when did it enter?
            • What’s the worldwide incidence of XMRV disease; that should say, “or prevalence” of XMRV. Where is it? Is it in England, in Europe, at what level? We know it’s 1.7% in Japan because of a study done earlier this summer.
            • And does it alter the risk of cancer development?  Because HIV & HTLV-1 both, by causing immune deficiencies, do.
    • [SLIDE 36:  WPI Collaborations International]
      • • A lot of people wanted to know: are we working internationally to replicate the studies? Everyone you see on this slide, a lady in Canada, part of the blood group in Canada also had called me since the study came out.  We’ve been working with Jonathan Kerr, and we have a 5-year RL1 with him. Ellie Barnes is in MRC in Oxford.  Norbert Bannert on that German paper, he was working with an advocacy group led by Regina Koch, I think, and they found a few samples that were maybe positive.  So he called me and said “Can we work together and have that antibody?”  Again everybody you see on this list, Jonas Blomberg in Sweden.  Norway, Germany, the Netherlands, Italy, Spain. We can’t handle the samples we’ve got so far, but we’ll try, and we’ll send the reagents out to anybody to replicate the work and find out more about the disease.
    • [SLIDE 37:  Additional Incidence Studies]
      • • We also know of additional incidence studies that we’re not involved with but are occurring at Kiel University, again in Germany.  And here in the USA, Sam Chow is working and has identified the virus in China, and I do know that Richard Huber has had success at finding virus in CFS and other patient groups.  And of course I mentioned that blood working group that’s working throughout the United States, and I didn’t list the number of US collaborators we have.  There is a lot of interest.  A lot of the world’s best labs are working on this, and we’re going to get there a lot more quickly than we would because of everything we have learned from HIV.
    • [SLIDE 38:  XMRV Clinical Diagnostic Tests]
      • • What about Diagnostic Tests?  Should your physician or you want to be tested currently (the last time I looked online), there were only 3 companies offering the diagnostic test. Of course the WPI licensed the technology to VIP Dx (we’re a non-profit institute), who is using our proprietary culture method and PCR, in combination along with a Western Confirmation. So, we look for both antibody and PCR positives in the cultured and co-cultured cells. And we use 20 mls of blood to do this, and make sure of the accuracy of the result, and the price is $450. Clongen lab has a real time PCR that is just looking for sequences on 1 ml of whole blood, and their price is here [$375]. You won’t find this virus in 1 ml whole blood by PCR. I think I’ve shown you that with the negative cases in the prostate cancer. And also a company known as Cooperative Diagnostics in South Carolina. We don’t know what their PCR method is, but they’re using a drop of blood on a piece of paper.  So, they tell you if you put a drop of blood on a piece of paper, you don’t need a doctor or anything, just send a check, and of course they won’t find anything.
    • [SLIDE 39:  Your Help is Critical to Advance the Science]
      • • Your help is critical to advance this science. At the WPI, we either want your money or your blood. That’s the only two choices! We need you to participate in these research studies, and we have a form online where you can register: https://www.wpinstitute.org/patient/volunteer_form.html, and you can email me.
        • We’re asking for some clinical characteristics, but we’re asking for those more to help us put it in a study.  We won’t turn anybody away. We will look for the virus, if we can get those studies. We’re waiting for the IRB approval, that’s the human assurance to make sure we’re not hurting you and we’re protecting your privacy. So, we expect that this week.
        • Donate funds to the WPI research and clinical programs that will be established later this year. The clinical programs will really come from the research and the diagnostics. And then write to your government officials, and encourage them to support XMRV research. This is an infectious disease. Why isn’t the National Institute of Allergy and Infectious Disease considering this virus? They’ve been pretty quiet, haven’t they? We haven’t heard a word from them. So, we need our government agencies to look at this virus because it’s an emerging infection, as I said, of unknown pathogenic potential.
    • [SLIDE 40:  Acknowledgements]
      • • I’d like to thank the people who – we couldn’t have done this study without them.  This has been a 3-way collaboration between the National Cancer Institute and its contractor SAIC, Cleveland Clinic, and the WPI.  As I said earlier, when Vinny Lombardi and I, together with Max Pfost, first saw the few sequences of the virus, I called Bob [Silverman] because obviously we were doing the work with him, and then I called [Dr. Francis William Ruscetti (born 1943)], and said, “I need you,” and he said, “I won’t go,” and I said, “I’ll pay your way to San Diego on the beach for a week!” and he said, “Not any more of your schemes, Judy!  I’m not going to do that again!” At any rate, we met at a restaurant, and we showed Bob, and he didn’t know what I was going to tell him.  Interesting, Frank’s a bit cantankerous because I gave him about a week’s notice because we’d had 3 weeks, and I was pretty sure I knew what we had.  We had to get a 3-way inter-institutional confidentiality agreement. Frank called the office in the government, and they said: “No, we’re not going to do that,” and he said, “Look, we’re talking next Saturday.  You can either have a confidentiality agreement or you won’t,” but they got one. So Frank and his lab, like Dan Bertolette, did every one of those beautiful Western’s that I showed you, just a magician.  Mike, Dean, and Burt Gold sequenced the entire RNAse-L gene in more than 100 patients. Ying Huang did all of the PCR that we had done, totally blinded, where samples never came to our labs to show it wasn’t contamination.
        • And, of course I’ve mentioned the lab of Sandy Ruscetti, Charlotte Hanson, and Jami Troxler, who were key in providing all of those reagents, without which this study clearly wouldn’t have been done.  Cari Petrow-Sadowski developed that immune-response assay in a real hurry this summer, and I mentioned Kunio & Rachel, who did bioinformatics support and electron micrographs. We couldn’t do it without Dan Peterson’s diagnostic skill.  I mean, he biased the patients such that we could find the needle in the haystack, but these are the patients that come to the institute.  They have classic symptoms of CFS.  When we have taken patients that have emailed us with exactly those same symptoms, we find the virus every time we look, including in Europe, in England, Ireland.  We couldn’t do it without the CFS patients and advocates.  We do so appreciate the support all along.
        • This was a tremendously difficult year in trying to keep quiet.  Knowing what you had and don’t say anything until you are sure you’re sure you’re sure.  Every day was just, “are we sure?”  And so, we were able, with the small crew you see here, and this supportive staff. Vinny, Katy and Max – they pretty well have worked 24/7 for at least the last year… So, at any rate, with that I’ll thank you for your attention and take questions.

QUESTIONS – Audience Q&A Session

• Question: What percentage of the population has this virus?
  • • Dr. Mikovits: We found it in 3.75 percent of the U.S. population, and it was from across the United States.  And in Japan they found it, just screening the blood supply of a couple thousand people, they found it in 1.7 percent.  So we don’t know the true prevalence.
• Question: So, only in 3.7 percent?  What about the other 96 percent of CFS?
  • • Dr. Mikovits:  No, that’s the healthy population.  It’s in 90-some percent of the CFS population.  I stuck this slide in here, and Frank keeps taking it out, but you might have heard in the press after the paper came out, we didn’t do all of the tests, all four tests, on all of the people prior to the submission in May.  We just looked for evidence of infection and looked to see if we could isolate the virus, looked to make the point, and it wasn’t so much about the CFS. What we did, after the paper was published, we went back and we looked with all four assays for evidence of XMRV in those PCR negatives.  Because now we know that indeed those negative samples may have evidence of infection, and what we found was that 19 of the 33 had antibodies in the plasma.  We found transmissible virus in the plasma of 33 of those people, and then we looked at that latent virus because the company I used to work at here in Santa Barbara was called Epigenics, and it was developing methylation-inhibitors for epigenetic silencing, and that’s what happens to viruses.  So we used Decitabine, which is a demethylating agent that opens up the genome and turns on the virus and found that there was latent virus in 10 of those people. And when we summed it all up and tabled it out, 99 of the 101 patients in the Science paper had evidence of XMRV infection. Another way that HIV/AIDS causality was established was by saying, “So, the statistics of this means there’s a 10 to the minus 35 chance that you had CFS in our study without having XMRV.” So I’ll go back to the AIDS analogy.  You can have HIV. So, we wrote in the paper “virtually impossible,” and the editor took it out. We wrote, “highly significant.”  The editor took it out. Finally we said, “significant.” But at any rate, 35 zero’s in front of a 1 would tell me it’s virtually impossible to have had CFS without having XMRV in this study. So, you can be infected with HIV and not have AIDS. We know that. People are being treated. A lot more elite-controllers are coming out, but you can’t have AIDS without having HIV. So if we can establish that XMRV is to CFS as HIV is to AIDS, which is what we’re trying to establish through that immune system understanding.  So HIV kills CD4 cells and leads to AIDS. XMRV does what to the immune response?  To the T, B and NK cells to lead to CFS?  To turn your question around, sir. What about – the incidence of CFS in this country is 1 to 2 million – is that more or less what is said?  By the Canadian or the Fukuda criteria?  So I said 10 million people were infected. Where are the other 8 million?  Do they have cancer?  Do they have nothing?  I showed you that only 20 percent of HTLV-1 infected people were actually sick with one of those diseases. So you can be infected with retroviruses and be carriers and not be sick.  And that’s one reason to be tested.  If there is a genetic susceptibility, which we’re looking to, maybe a reason, an immune defect that was unknown as to why some people get sick and others don’t.  You certainly want to know where the virus is, so if you’re a carrier you can protect your family.
• Question: Do you know how many have tested with VIP-Dx, and how many are positive?
  • • Dr. Mikovits:  I don’t work with the company. They only take samples two days a week because it takes three days to do that, so they’ve done hundreds of samples in the last couple of months, and at least half of them are positive. Or 40 percent. And again, their doctors are looking at, the doctors who are well versed with CFS, so they’re immediately sending off. Dr. Cheney, Dr. Klimas, a doctor in Canada, Ellie Stein, maybe even Susan Levine in New York. I’m not sure, because it’s illegal for me to know those data because there’s confidentiality between the patient and the physician. But quite a number and, yes, it’s there. CFS is a heterogeneous disease. I mean anything based on fatigue. So certainly everything is not going to be this virus. But maybe there is a disease, and of course that’s what we’re looking for, biomarkers for understanding how you can get sick and be sick forever and not have drug targets, not have diagnostics.  Certainly there are going to be lots of people who have what might be called CFS today, and that’s why we’ve also coined the term XAND, for XMRV-Associated Neuroimmune Disease, and that would be because we’ve seen (inaudible). I had done a number of studies with family members after the paper came out and prior to now, where I just said there’s a family member where the children have autism, there’s fibromyalgia, there’s excess cancer, and when you look, you find the virus. So, we’ve found the virus in Atypical MS. Atypical MS is a non-demyelinating MS – it looks like MS, it has some brain lesions like MS on SPECT scans.  At any rate, I’m not a physician. Just looking at families with different types of neuroimmune disease, we started seeing that the virus was there, and so that’s why we started thinking it might be involved in a broader spectrum of neuroimmune diseases, with overlapping symptoms because in fibromyalgia, pain is the primary symptom, but, in a lot of people, it’s this body-numbing fatigue, so fatigue goes along with it. In fact, Cindy Bateman, who’s a fibromyalgia expert at the University of Utah, says fibromyalgia is CFS with pain, and she can distinguish those who get better with fibro with certain therapies and compounds, and she takes the others away who don’t respond at all, and puts them more into the CFS group. So definitely XMRV is not going to be everywhere.  It’s not – it’s 4%, but the people who are infected are sick.
• Annette Whittemore: Earlier you said that 40% were positive. So describe the fact that if you’re positive, you’re positive.  But if you’re negative, you’re not necessarily so?
  • • Dr. Mikovits:  Yes, that’s correct.  I answered that question based on the samples that came through there. Everyone who is positive is definitely positive for having the virus. But we don’t know what the people are, what the doctor is sending in, so the people might not have that disease. So it could be a clearly, distinguishing delineating marker – biomarker – or diagnostic at that point for various diseases. So a doctor might see a spectrum and say “I don’t know, maybe I’d better check.” Because the earlier you catch it, just like cancer. Early detection. Make sure the reservoir is (inaudible), make sure you don’t have that virus multiplying, and you can live a normal life. Don’t let it get (inaudible). You know the commercial out right now is “HIV doesn’t have to equal AIDS”; well XMRV doesn’t have to equal disease. If we keep it down, we keep the immune system strong.
• Question: So what you’re saying is you may test negative but not be negative?
  • • Dr. Mikovits:  That’s correct.  If you do it by the PCR.  If you do it by VIP-Dx, at least right now, it’s running along the lines of (inaudible). We’ve got the antibody, and we’ve got three of the four tests. We’ll license it to anyone. We’re a non-profit institute, so everybody pays the same royalty, so any diagnostic company could do the gold standard. But right now, if you test negative, you’re not necessarily negative, even at VIP-Dx. Because we want to go do that serology test. Maybe we can’t find evidence of the virus.  But you’ve been exposed, which would be a good thing because your levels are theoretically low, and you’ve just now made the antibodies, so you can prevent disease, as we did with Magic Johnson. But we don’t know anything about the immune response to the virus.
• Question: What about transmitting it, when you get pregnant, to a child?  I mean, you were talking about breast feeding?
  • • Dr. Mikovits:   Well, it is theoretically possible. We don’t know. Gamma retroviruses are vertically transmitted, so the egg and the sperm can be infected and you can actually vertically transmit gamma retroviruses. But this is the first (inaudible), we’ve only studied it for two months. We don’t know.  Theoretically it’s possible. If we make those data, it will certainly fall out when we start looking at family studies. The horizontal transmission is the only thing we know about right now. Because, of course, you have to find somebody who just gets infected in order to understand the disease.  And since a diagnosis of CFS is being sick for six months. Well, if that’s the only thing we can do is to stop that practice, making somebody wreck their immune system, or be totally sick for six months, that would be a great thing with the discovery. If that’s the only thing that comes of it.
• Question: Would you not recommend getting the test again if you did [test negative]? I just took the blood test with my doctor… and he’s part of a study. He didn’t tell me who he sent the samples to, it’s a 20-patient study, so if that came back negative …
  • • Dr. Mikovits:  It depends on where it is sent.
• Question: Take, for example, VIP-Dx?
  • • Dr. Mikovits: If it’s at VIP-Dx, as soon as we have the serology, we’ll go back and do all the negatives.  We save them…  We do isolate virus from all of them, but it costs a lot. So if they’re really negative, we won’t isolate virus either. That’s correct.  At VIP-Dx, we’re going all the way to virus isolation, because we want to make sure we are sure.
• Question: So, at a non VIP-Dx?
  • • Dr. Mikovits:  They’re only doing PCR. All bets are off.  I showed you almost 60% of them are probably, they’re all false negatives, because very few people can find them. And participate in our research studies, because as I said, there’s no hurry. There’s no real reason to pay for a diagnostic test because we can participate in research studies and fund those studies and you get better data. Because I’m not privy to those data. I can’t answer the question of where they came from, where they are in the United States.  Those patients and doctors have to release that information, and that’s not the job of the diagnostic company.  So the information is not usable in that way. We’re pretty good right now. We’ve been as responsive as can be. By the way, if anybody thinks they might have been in our repository, if you simply e-mail me, we did decode that study over the Christmas holidays, so we can tell you if you’re positive or not.  We can tell you whether or not you were in the study.  Because there’s more than 500 samples, who are patients in the repository, and we only pulled about a hundred of those for this study.  I don’t remember everybody, but we did decode it, so we know who the positives are at this point, and we are sending letters, but you have to ask me first for our human-assurance protocol, so just ask me and we’ll tell you.
• Question: In the beginning, you mentioned that the virus uses lipids to grow?
  • • Dr. Mikovits:  It actually uses the cell lipids to make its lipid membrane. It’s an envelope virus, and it has lipids, so it pulls cholesterol in to make its (inaudible), so it uses all the cell machinery. It only codes the enzymes and the envelope proteins and cells, so the lipid bilayer of the envelope virus, there’s lipids in it.  It’s from your cells, and cholesterol’s a part of that.
• Question: Because I’m wondering, I’ve met a lot of other CFS patients who like me have high cholesterol. That just made me think: might there be a correlation?
  • • Dr. Mikovits:  Yeah, and nobody’s ever looked. It’s certainly something that they could look at correlating. I can’t think of a reason why. You might presume you’d have less, if you’re using it up for another purpose.
• Question: What about children?
  • • Dr. Mikovits:  We do have a little bit of data on that because we have two children in a study who have a genetic disease of cholesterol.  It’s called Niemann-Pick’s Disease. It’s also known as Childhood Alzheimer’s. And these kids, you know it’s a cholesterol metabolism disease where eventually your brain, you’ll eventually die of it because if you get too much cholesterol, it messes up your brain and everything. And those kids have been treated by James Hildreth in Nashville, Tennessee, at a small college…  He’s using Cyclodextran and some of the cholesterol drugs.  He’s actually an HIV drug developer, and the kids are showing some improvement when he modulates that pathway and stops the virus from entering or exiting the cell, so we don’t know anything about XMRV.  We just know what other viruses do, so he is having success, suggesting that there are some opportunities there.
• Question: Are you working with him?
  • • Dr. Mikovits:  Yeah, we’re working with him as well. That’s why I didn’t list all the collaborators in the United States. We’re providing reagents and whatever intellectual knowledge we have, and whatever physical abilities and instruments we have, to these collaborative efforts as well.
• Question: Until forthcoming therapies are established for those who have XMRV, what are people doing once they are tested?  Are they taking immuno-modulating therapies or whatever?
  • • Dr. Mikovits:  Some of what I showed you here, that turn on/off switch, suggests non-steroidal anti-inflammatories.  So non-steroidal anti-inflammatories could help.  Things that will balance cortisol.  Maybe – these are just thought processes – because you know inflammation turns on the virus, and I don’t know much about hormone therapies and how lowering hormone levels might help.  But I do know anecdotally that a lot of women in a particular time in their cycle get much, much sicker and can’t get over it. I do know in the laboratory progesterone really up-regulates the virus, so if you have a birth control pill – and again I don’t really know anything about this (I’m not a physician) – you might think about just keeping the levels balanced and avoiding the fluctuation. So certainly supplements can help a lot.  Retroviruses cause a lot of oxidative stress.  So things like N-acetyl-cysteine and glutathione, the detox type.  People do take supplements.  I know that a lot of people have had success with immune modulators, just helping their disease, or supplements, because they know about them.  I caution against taking too much or taking a bunch of things. Try to learn as much as you can, because supplements aren’t controlled by regulatory agencies, and therefore, if you’re not using a high quality, you could actually be putting poisons in you.  Since we don’t know much about the virus, you could modulate the wrong way. But things that up-regulate Natural Killer (NK) cell function (and there are known compounds out there that do that, that are marketed in our state), so that could help you.  So, we don’t know a lot about it, but that’s how people are actually starting to help themselves. The other thing is to stay out of stressful situations. It’s hard to do. In fact, we’ve seen a lot of people get worse just with the stress of the discovery, which is sad. That’s why I want you to call me. Because we don’t want you to think: “Oh no, I have a retrovirus!” We want to talk to you because it’s serious and you can have untold (inaudible). I mean most people say: “Wow I got it!” you know they’re congratulating people when they come up positive, which is really strange…  [laughter]  Then they get really scared, because they don’t know anything about it. And we’re here as much as we can to help, but we don’t know anything about that retrovirus. All we can say is the same thing I’ve been saying today. I hope you would walk home and say: “It’s not a mouse retrovirus.  Retroviruses are not ubiquitous, and they’re not benign. So I have to think about those facts.”  It’s wide open. The drug companies – the one thing, if you do get tested, and we know you’re positive, a confirmed positive – we’ll get you into the earliest clinical trials.  And they’ll have things pretty soon because it’s major Pharma, and I’ve given them the reagents and the cell lines – we’ve made several cell lines from people that make a lot of virus. And so we’ve given them those cell lines. All they have to do is take something off the shelf that rationally might inhibit a particular path, say the integrase gene or another gene that’s conserved across the three retroviruses [HIV, HTLV, XMRV] and show that they can see the efficiency, efficacy and knock down levels of the virus in the lab to levels, which they’re the known blood levels of the drug can achieve, and they can submit and do the paperwork for a clinical trial. And it’s already known to be safe because it’s already passed Phase 1, or safety trials, in humans. So we looked at those first. And there are a number of companies who, as I say, they are high quality companies, and they are more than interested. They are doing it now, and have been doing it since October.
• Question: What about cancer, especially the hormone-responsive cancers?
  • • Dr. Mikovits:  We very much expect that some of the breast cancer incidence – we hypothesize that inflammatory breast cancer is a lot like what we saw with the inflammatory prostate.  But yes, it is a very real hypothesis because the incidence of breast cancer in young women that you never used to see before, is rising at levels that suggest something environmental, and not necessarily genetic.  You know, “we never had cancer in my family,” and you see young women that way, so it certainly is something that we’re looking at.  The “we” here – I always say “we,” and it’s everybody but me usually – it’s the National Cancer Institute.  We’re also looking at lymphoma, because CLL (chronic lymphocytic lymphoma) is a lymphoma, a B-cell lymphoma and it’s also been going up and up, and it suggests to us some kind of role of an infectious nature. So we are looking at a number of lymphomas with a group in New York, a group in Florida, and the Nevada Cancer Institute.  I don’t have a breast cancer study set up.
• Question: There’s actually an anti-retroviral vaccine that’s been used…
  • • Dr. Mikovits: Yeah, a vaccine is a real opportunity, and we know that they still don’t have an HIV vaccine yet that’s efficacious, but HIV is a complex retrovirus. So when you’re thinking about the reason why you have to take a flu vaccine every year, it’s because the virus changes. Well, an HIV virus in a person in a week will change too much.  They call them quasi-species. One of the really interesting things about this study is that we only isolate one thing out of these people. When we do the sequencing, it’s clean. We don’t isolate quasi-species. We don’t have the virus have these changes here in one week or one year – we have patient samples across dozens of years. We isolated XMRV from a 1984 plasma sample from a patient. We got it in 2008 and we got it in 1984, which again suggests that the virus has been around at least 25 years, and it might have a role in the disease but is not causative. So yes indeed, it could play a role in other things.
• Question: People with CFS have shown a lot of evidence in other studies of being immune-suppressed.  How are you certain that it’s XMRV that’s causing immune-suppression as opposed to say 8 other viruses like Kerr is suggesting, or other viruses that are causing immune suppression and making XMRV opportunistic?
  • • Dr. Mikovits:  Well, there are a couple of things for that.  First of all, we’re not certain of anything.  So I’d say it’s a hypothesis.  It’s because of what I know about HIV, and HHV-8, herpes viruses, where it’s the underlying immune deficiency.  The other viruses aren’t retroviruses, the other pathogens too, the bacteria, and they don’t live in your immune systems forever and replicate and have reservoirs.  They’re across the board, so everybody’s infected.  Probably 90% of this room has an EBV infection.  But very few people express EBV, have chronic active EBV.  That suggests that your immune system has something wrong with it.  It could certainly go either way, but retroviruses don’t do that. The CFS world has looked at many of those pathogens, so here’s chronic Lyme and here’s EBV and here’s…  It’s never one place, something that unifies all of those.  So it’s certainly a testable hypothesis, and that’s one of those things that will just happen.  If you get an anti-retroviral and the chronic EBV goes away, and a lot of the symptoms go away…  I’m not saying that the EBV doesn’t cause a lot of those symptoms.  That’s what makes it so hard to figure out the disease.  But if there’s an underlying immune deficiency that’s created… that’s not simply depression… but is getting worse every year, could be an explanation.  So we’re happy that we can test that, because we do have different populations where we can see what the role of the co-infection is.  We’ve never looked. We’re looking with various groups at big cohorts of chronic Lyme and big cohorts of chronic EBV, Q Fever – things like that have been associated with.  Jonathan Kerr, in fact, he’s working actively with us to see if it makes sense, that you need the combination or you need one or the other.  But, in the general population, the incidence of XMRV is something between 2% and 4% right now, whereas it’s 90% of some of the herpes viruses.  And most of us are exposed to some of these other pathogens, so I certainly don’t have an answer, but again, this gives us a testable hypothesis to look.
• Question:  I’m XMRV positive. And I have a son who is. I was told that he also had Lyme when he was 7.  Explain the chronic Lyme connection in him?
  • • Dr. Mikovits:  Well, again, we don’t know – it’s our hypothesis that, especially in Lyme disease, where it goes away and it’s almost cured and you only see some proteins that don’t necessarily; you know; it suggests you almost cure it with the antibiotics, but you have to keep the antibiotic there because there is a low level that your immune system can’t clear, and maybe it can’t clear it because you’ve created an immune deficiency with the retroviral infection. We’ve never looked at a Lyme cohort yet. Again, we’re setting that up, but we don’t know the connection. The hypothesis is, if we can treat the retrovirus, then the chronic Lyme will go away.  And you’ll treat both.
• Question: But he was untreated until about 7, and was bitten in Europe, and no-one understood that there.  He had the rash on his legs, and no one believed me that that’s what it was
  • • Dr. Mikovits:  Well, we can still clear the Lyme. For instance, in the AIDS population, you treated the pneumocystis pneumonia. You treat it with the appropriate antibiotics because you don’t want the co-infections to kill him, and then do the anti-retrovirals too. There’s no reason, for instance, one of the questions that I got online was, “Well, I’m taking antivirals. Do I need to stop in order to get tested?” No, because antivirals don’t target retroviruses. Retroviruses are very distinct viruses, so no you don’t need to stop. We’ll still find the virus.
• Question: But even 10 years later, because he was never treated. That was 10 years ago. You’re still saying you would still treat for Lyme?
  • • Dr. Mikovits: Yeah, well you probably should be at this point. Treated for both the Lyme and once we have a treatment, for the retrovirus.
• Question: If XMRV is transmitted sexually, how come it’s not seen as CFS in couples?
  • • Dr. Mikovits:  The possibility is that it’s transmitted sexually, but we’ve never actually shown human-to-human transmission, meaning we caught the day when the other got sick. I don’t actually have an answer for that, other than that I know that it might well be more in couples than we think, because it’s a milder form of symptoms, or maybe this person’s a carrier. There’s still a lot we don’t know about why prostate cancer and why CFS… What is the hormone component that turns on the virus? They may be carriers and not know it, and certainly there’s a lot to study there, to understand the gender differences in these diseases.
• Annette Whittemore: Maybe part of the answer is that, if XMRV turns out to be the cause, would you have said that you wouldn’t have CFS without it, given you can have the virus without having CFS?
  • • Dr. Mikovits:  We have looked in a limited number of families that we’ve done, and in fact, maybe only one member of the family has CFS, and so it will be an interesting year, but, once we really get to the data of looking at these families and the various diseases and trying to understand – it’s a very low replicating virus, so it really just sits there for a long time. If you can keep the reservoirs low, you might have the virus your whole life and never get sick. We don’t know how long it’s been in the population. We think, like with other retroviruses, the younger you get sick, the more severe the disease. When Sandy Ruscetti gives the rats the viruses when they’re neonates, they get cancers. When they get it when they’re 30 or 40, they don’t get anything. The immune system is educated and grows as you go along. It can be more fragile at different times in your life. We’ve seen a lot of puberty, boys and girls alike. There’s a lot of infection or at least apparent infection, disease occurrence at 12 or 13. And I do know that’s when Andrea got sick [to Annette Whittemore]. And yet presumably, your family was in the same space, but there was no child who had hit puberty at that point, and that might have been the difference. Retroviruses don’t infect people differently.  You can’t go to Germany and say the reason they don’t find it is because they have hardier genes in Germany (although my husband might argue that) [laughter]. Because everybody gets infected.  It’s just which immune system can control the virus and keep it down. Since we’ve been able to treat HIV/AIDS now – we found elite controllers, people who are walking around with HIV who never knew they had HIV, and the copy numbers are all low.  Their immune system is fine, and they have no idea when they got it.
• Question: A lot of patients say they had a flu, a weird flu-like condition, a week or two before they got CFS. What you’re saying is that flu-like illness is a bug that came along and allowed the XMRV to create CFS?
  • • Dr. Mikovits: Well, no. I know almost nothing about CFS, but what we think is happening is – remember the slide where I showed you with those little events?  You know, what was the event that was the straw that broke the camel’s back? Where did the balance tip between here you’ve got your immune system working well and the virus and the immune system are co-existing just fine, then some other, that other bug, whether it be Lyme, a flu, anything, gets you. Then the virus, the cells divide, and so do the B and T cells you need to mount an immune response. And now you’ve got your memory population that might have been harboring the virus, and it’s replicating because it’s seen that same pathogen before, so it could be a common everyday pathogen, and then you just tip the scale to where now your immune system can’t handle it. And every day you’re seeing more infection because your NK cells aren’t working, your B cells aren’t working. We put that antibody up there for a reason. We haven’t been able to correlate the levels yet, because we haven’t been able to find high enough numbers. But we do see these infected families where infected spouses have very high levels of antibodies, that suggest maybe antibodies in this retrovirus can be protective, and maybe there’s an immune therapy on the horizon as well. So you can think about it in that way.  It doesn’t have to be the insult.  You might not know how long you harbored that virus.
• Question: So what you’re saying is XMRV was there, it wasn’t the insult, then something else came along that was the tipping point?
  • • Dr. Mikovits:  Yes, that was the tipping point, that’s correct. That’s our hypothesis. And again, it’s testable, we don’t know.
• Question: You’ve got a big population of patients with sudden onset and then you’ve got a big population that had gradual onset. And a lot of the gradual onset patients are worried that “Maybe I don’t fit this equation.” What would you say to that?
  • • Dr. Mikovits: That little bump is smaller, so it’s not a huge burst. That it was little insults over time. I think, for me anyway, I know only a handful that I’ve looked at, that I know the patients, and I know what the onset was. But the gradual onset – there’s no real difference. It just depends on that environment, what the other triggers and events were that spurred it on. So, I don’t see, if you are gradual onset, it doesn’t mean that you might not be infected as well. Questioner: That’s an important thing to know.
• Question: Have you tracked any of the inflammatory markers in the blood with the XMRV virus?
  • • Dr. Mikovits:  Yes.
• Question:  What are you finding?
  • • Dr. Mikovits:  We find a signature suggesting a viral infection, an unclear viral infection.  We have 5-10 inflammatory cytokines and chemokines that will cluster in an infected person. The problem is we don’t know if it indicates active infection. Certainly, when the virus is quiet, that inflammation will go down, and those cytokines will change. And it might be a nice biomarker for following active infection, but we haven’t analyzed the data in that way yet. It very well could sit, and some of those go down and up very quickly.  So, like in 3 days, your IL-8 and your IL-6 and some of the chemokines are elevated and then they go back, like an EEG. Just like you might assume the retrovirus could be an EEG, depending on where in the body that is. We don’t know the reservoirs, so we don’t know what’s controlling it.
• Question: I’m confused about something. You say you can’t find it very easily by PCR, so you are culturing it. Then how did you find it for the Science paper?
  • • Dr. Mikovits: Well, because I found it in 67%, by PCR, of the patients, but I looked at samples collected at several times. I just said it’s like an EEG. So I got lucky and I found it at a time when the patient was high. They’ll come to the doctor when they’re sick, which might mean they’re replicating more white blood cells, and there’s more virus in their white blood cells.
• Question: So you took like 10 samples from one patient?
  • • Dr. Mikovits: Not that many, usually it was 4. The copy number could be as low as 5 or 10 copies per mil of blood, so there’s a statistic called a Poisson distribution, where you might find it one out of three times, so we went more than one of that and went 4 on most people when we found it.
• Question: So the UK paper had studied patients, if they had looked at 4 or 5 per patient, do you think they might have found it?
  • • Dr. Mikovits: They might have certainly found more, yeah.
• Question: They essentially accused you of contamination.
  • • Dr. Mikovits: Right. But why would I have contamination in my sick people and not my healthy people?  How would I do that? I did it in three different labs. I did it in Cleveland Clinic. When I first came to the Institute, we didn’t have a lab. So everything we drew from around the world, I sent to Frank’s office.  They processed it. I went there, I put it in the microarray or did whatever I was going to do, and we worked there for a couple of weeks, you know, while we were building our lab and it got started. From the way we did the study, there’s just NO possible way there was contamination. And that’s what the reviewers concluded. And the phylogenetic analysis was one of the things they asked for (they asked for 3 things after the initial submission). The phylogenetic analysis proved two things. It wasn’t a mouse virus, it’s a human virus; it wasn’t a contaminant from a lab. We never do mouse work, but it wasn’t a contaminant from mouse feces or something in the lab, and it clearly was a new branch in the [phylogenetic] tree, a human virus. And our virus wasn’t exactly the same as the prostate cancer virus. It’s still XMRV, because it’s 99% similar. But that’s enough to show it’s not a contaminant. One of the things you have to use to get a good PCR is at least 750 nanograms of DNA. They have no idea how much DNA was there. And they quantitated 3-9 out of 186? Sure they found a band of globins, but globins are in every single cell, so again you’re making an unfair comparison of what you’re saying you see. And then you amplify it for fewer cycles than what would really push the envelope. We’ve also done – to show no mouse contamination with the CDC, and Bill Switzer – after he saw the results in the paper before it became published. He said, “I have an assay that’ll show it’s a mouse contamination. It’s a very sensitive, very specific PCR. Will you do it?”  I said, “Sure, send it to me.” We did it on all 100 and not a one. Not one cell line in our lab. He’s found it in a couple in his lab, but we didn’t find any. Perfectly controlled. He said, “Congratulations, it’s not a mouse contaminant.” So there’s little else we can do, except wait for the rest of the community.  It’s there. And the prostate cancer people didn’t say: “oh you didn’t find it in 500 people; it must not have anything to do with prostate cancer.” Because it just didn’t have anything to do with that population. And again, Norbert Bannert is a high-quality scientist. As soon as he saw the paper, he called me and asked for the reagents. Because he’s gonna go back and look to see if it really is there, and help us find some answers. He’s also looking at a CFS group. So it just depends on what you really want to find. We weren’t biased in our study. You know, I’m a cancer cell biologist, and we aren’t biased. I had to work really hard to get most of the people – not Frank and company – the National Cancer Institute (NCI) didn’t know what CFS was. You know, fortunately, our scientists can to some level do what they want, as long as it’s along with the mission. I remember one gentleman, high level NCI official, said “tell ‘em to get over it.”  Again that’s the credit to Annette and the formation of the WPI – we got a grant where we literally named XMRV within my first six months in 2007, because of the juxtaposition of seeing that paper in prostate cancer, right when we met. And thinking about the possible mechanisms, and the grant got rejected three times, you know, because scientifically, retroviruses aren’t in CFS. If you went to Wikipedia in August, it said retroviruses aren’t in CFS. It doesn’t say that anymore.  So we made progress.  [Audience: Wow!]  So, will there be a variant?  Maybe England is the variant. Maybe there’s XMRV2, and one causes more.  Maybe we found the one that causes the least severe disease, and maybe there’s XMRV2. A group emailed me from China, and the guy said “please,” you know in very broken English, “please don’t leave me, please write me back. Please help me.” And so Sam Chow was going over there, and I said will you look up this group? And Sam Chow has found a virus that looks like it might not necessarily be the same virus, might be a lot more aggressive virus over there in disease, but that’s just anecdotal. So, we don’t know. The good news is we have something to work with that is a very testable hypothesis.  This has been very rigorous. You don’t get more rigorous than Science, and certainly not Frank and Sandy Ruscetti. They’re highly regarded. That entire team contains world experts. So, the CFS population had the opportunity to have them look at us, and they are good enough scientists that they didn’t show any bias.  They looked, you know – I asked Frank to come out to Reno before I took the job, because I said, I hear a lot of things about this. So he came out and stayed a few days and talked to some patients and spent some time with them. And I was on an East Coast trip working with my drug company.  I came back a few days later, and I said, “Well, what do I do?” And he said, “Take the damn job!” So, if you’re really looking unbiased, and you look at this, it could be only 10 million people in America, it could be only endemic right here, it could be like Japan. And HTLV is pretty-well innocuous. 5% of people get ATL and 20% get that tropical spastic paraparesis or HTLV-1-associated myelopathy. I was glad when they changed that thing, by the way, because I can’t say that and I couldn’t spell it either! Those patients, they actually go on steroids to dampen the inflammation, the immune response, and they are fine for decades. But you know, in the Caribbean and in Japan, it was a health problem, so maybe we only have a health problem and the distribution is not – but we do have at least 10 million Americans, and maybe a majority of the CFS population here in America, that we have a lot of work, just to treat, and that will be the focus of the Institute in the coming years. Certainly, we will treat everybody else, but obviously we don’t see anyone else. Dan was funny, he was like “Well, Judy, I have to take care of everybody else” and I said “of course, there’s not anybody else!” So it’s an interesting and an exciting time for sure.
• Question: I just wanted to say, you know, it is really exciting to hear you say we need to do this, we’re gonna do this. The virus you are studying, it’s our virus, and I think it is really important to note everyone that for you to do what you’re talking about, and for other researches to do it, we patients need to get behind funding the research in a way we’ve never done before, or it is not going to happen.
  • • Dr. Mikovits: But have your government fund the research. HIV incidence in this country is about 800,000 people, I just quoted 10 million….. I shouldn’t say this, but when I saw it was a prostate cancer virus, I said “man, those men can’t possibly ignore this now!”  [laughter].  I never said that publicly.
• Question: Would you talk about what is coming up in the next years, what are the next steps, where are possibilities with your treatments, with the replication studies?
  • • Dr. Mikovits: Yeah, all of those people I showed you internationally are working to replicate the study as is the Blood Working Group. We have been intensely working with them. We have another conference call on Monday, these things are happening. We will put probably 20% effort in our lab into that study. We are very serious about transmission studies in our lab and in the Institute, and we have one starting where we’re simply comparing infected people, people who we isolated from the blood, and just taking DNA in their saliva, for example, to see if there is any evidence in saliva or of that kind of transmission just again. That is just because of anecdotal stories, where people say, well, you know a bunch of kids on the playground with a water fountain, or at school. I don’t know what the anecdotes are, but just thinking about ruling those kinds of things out is a study we are doing. We are also actively looking at the incidence in other neuro-immune diseases, so we are looking at that study I told you about.  We are looking at cancer and CFS, we are looking at Fibromyalgia, Atypical MS. We’ve got a study going with Vanderbilt in POTS, which is Postural Tachycardia, because of the overlapping symptoms.  He is simply just sending me a bunch of samples, both sera and DNA, and we are just going to take a look to see if we see it there. Autism – we do have families with some autism, and there are some immune defects characterized by Judy Van de Water at the Mind Institute in Sacramento, and she sees some NK cell dysfunction, some inflammation, some of the things I told you about there. So, in that group of autism, we are looking to see if maybe there is not an underlying pathogen or XMRV infection.  Those are just the priorities, just in the coming year. The NCI, the National Cancer Institute, has already put $1 million into the development of the reagents and the assays, so very soon the best of tests and all the reagents can be distributed. So, the AIDS reference program (there is an AIDS Reagent Reference Program that if you just google that you will find them) has agreed to set up for XMRV and send reagents around the world. We’re spending considerable time and resources just shipping…We are continuing the studies of the immune system, so I had an entire program set up from the beginning where we are looking at the genetics with Mary Carrington. We are looking at the type 1, so Vinny Lombardi will continue his studies. Just because that single nucleotide variant, we didn’t find having any correlation with XMRV infection, doesn’t mean there’s still not something wrong with RNase-L, and that might be a therapeutic target. So Vinny is actively studying research, the type 1 interferon pathway and RNase-L. Isabel Barao-Silvestre is a faculty member that’s just joined us, she is a professor, and she is at UNR as well, and she is an expert in Natural Killer cells and killer cell function, so she is doing a lot of the innate immune response and understanding how XMRV infection in NK cells might contribute to disease. As I hypothesize, we don’t know how yet, so those are the internal programs going on. Because of my background I’m actively perusing all the drug development efforts in our laboratory by working with at least three companies right now, to look at that. Those of you in the audience and around the world, if you email a question, we try to answer every one of them (info@wpinstitute.org). One that came up which I think is good to address is, a lot of people wanted to know if XMRV somehow “piggybacked” on EBV or other pathogens to get into an affected individual, and I didn’t know what that term meant. It’s not a scientific term. But if it meant that you couldn’t get XMRV unless you had come in with another infection, there is just no evidence of that in any retrovirus. So, yes, people think that people got infected with HHV-8 and HIV at the same time because of the Africa thing. You can get infected with 2 pathogens at once, but there is no need for any to piggyback. You don’t need another pathogen in order to be infected with XMRV or any other retrovirus. And any other question, if you have written them down, I am happy to answer them. Some people are like “I emailed you yesterday and you didn’t answer” and I say “oh, I slept yesterday.” If I don’t answer you within a week, then write me back because sometimes I miss it, and sometimes our emails are so full these days, that they are throwing things into spam. So, if you haven’t heard from me, you will usually hear from me within a week because I really do try and answer essentially every one that I get, which is probably foolish, but I like to actually, I like to work with the patients.

• [Andrew Bokelman] Q: From what I read, XMRV is infecting immune cells. If patients take nutriceuticals that have been shown to increase natural killer cell count, could some of this increase in NK cells come from dividing infected cells, thus risking a heavier load of XMRV?
  • • A: Because XMRV research is in its infancy, little is known about the tissue reservoirs of the virus, while many cell types can be infected (because the XPR1 entry molecule/receptor is on every cell in the body), few cells support high levels of replication of XMRV. You could potentially increase the viral load by increasing the NK cell count. However, we have done studies with compounds that increase NK cell Killing without increasing NK replication or XMRV viral loads so NK cells kill better and can therefore be efficacious in killing XMRV infected cell.
    • An extremely minor species of HIV becomes a dominant species when a large mixture of the viruses (quasi-species) from the blood of AIDS patients is cultured in the laboratory. As much as 18% of our human genome (3 billion genetic codes) are made of retrovirus sequences. The sequence of XMRV is 95% homologous to the human endogenous retrovirus (HERV) sequence in our human genome. Is the 5% genetic sequence variation enough to call XMRV a new virus? Is XMRV a minor quasispecies that is selected out and predominantly amplified from the tissue cultures?
    • The 6 full length XMRV clones: 3 from prostate cancer tissue biopsies from men in Cleveland and 3 from leukocytes from female CFS patients in the Western United States drawn in 2006-2008..were 99% similar. We did not despite repeated attempts isolate ANY other variants from those individuals suggesting that the virus is very low replicating and unlike HIV does not have quasi species. As much at 18% of the human genome is retroviral elements or endogenous retroviruses. THESE ARE QUITE DISTINCT FROM EXOGENOUS VIRUSES! PRIOR TO THE SCIENCE PAPER THERE were ONLY 2 EXOGENOUS human retrovirus families, HTLV (1,2) and HIV (1,2). HERVS are not infectious exogenous retroviruses...The main point of the science paper was the demonstration that XMRV was the third human exogenous retrovirus and that it was blood borne and transmissable (can isolate it and do primary and secondary infections of primary human cells) Moreover, the full sequence of XMRV was blasted against the entire human and mouse genome and NEITHER the mouse genome or the HUMAN genome contained XMRV..proving it was NOT a mouse virus. XMRV was on the basis of our data proven to be a NEW HUMAN Blood borne infectious and transmissable retrovirus. Finally the proof of an immune response in CFS patients from whom the virus had been isolated and sequenced showed that the XMRV in CFS patient was not a mouse contaminant or a human lab contaminant.
• [John K. Chia M.D.] Q: It has been said in the new paper article that XMRV is transmitted by blood, sex and body fluid. The author may have meant that this is what happens in mice but is not yet defined for human. Sexual transmission (men who have sex with men) and transmission through blood (hemophiliacs) were long suspected based on epidemiologic data before HIV was found to be cause of AIDS in 1984. ME/CFS does not have typical pattern of sexual transmission although there are rare mentions of transmission between couples. Most of my patients have developed ME/CFS after flu-like illness with respiratory and/or gastrointestinal symptoms, which would speak against sexual and blood transmission. Furthermore, why are children developing this illness and why clusters of cases occurred in Incline village and other areas, if the transmission is as stated. Please clarify this issue.Thank you,
  • • A: Newspapers are notorious for extrapolating data and drawing conclusions. We and Dr. Stuart Legrice attempted several times in writing to correct the mis-statements in the Wall Street journal and the NY Times. The Science paper showed that XMRV was blood borne, infectious and transmissable SUGGESTING blood and body fluid transmission although NO studies have proven transmission of any kind. The authors meant this is the mode of transmission for the other two human exogenous retroviruses HTLV-1 and HIV. XMRV is NOT a Mouse virus and NO ONE knows how it got into the human population). This author has never worked with mice.
    • It is important NOT to think only of HIV when considering modes of transmission. Body fluid means ANY body fluid, blood was proven in the Science paper, but one can also consider saliva, vomit, urine and feces. XMRV is a gammaretrovirus (simple) and these tend to be much more stable than HIV and HTLV1 which are complex retroviruses. XMRV has been demonstrated by us in the science paper to be very stable and easily transmissable CELL Free unlike its cousins HTLV (which is tightly cell associated) and HIV, which is very labile and not stable in vomit feces or urine and while it can be shed and theoretically transmitted in saliva this is rare as it is labile cell free as well.. the flu like illness and gastrointestinal issues are consistent with stability in other body fluids and particularly in acidic environments such as the gut and urine...We have isolated infectious XMRV from saliva and prostatic secretions to date and tested its stability. These are unpublished data and we await the development of a quantitative viral load assay, but when we can quantify the stability, my hypothesis is that XMRV is the most stable human retrovirus to date and while no human retrovirus (or animal retrovirus is transmitted airborne) our hypothesis is that we will find the transmission is in other body fluids and outbreaks occur because of the stability of XMRV in other body fluids 
• [Pat Turello] Q: Do you think that any of the new biotech drugs for MS could be useful for CFIDS? Thank you.
  • • A: My (limited knowledge of the new MS drug is that they target the hyperactive immune response and trafficking of white blood cells from lymph nodes so if a reservoir of XMRV is the lymph nodes and the hyperactive immune response (as we suspect) plays a role in disease progression through increased replication of XMRV or trafficking of white blood cells harboring XMRV to sites of inflammation, then these drugs could be useful for XMRV associated CFIDS. Are there any official ( or unofficial) records of forested areas near and around Incline Village being sprayed for 'insect control' prior to the outbreak of CFS there ?There is no data or records that I am aware of and no evidence that XMRV is spread through insects (in fact there is no evidence of how XMRV got into the human population)
• Q: Is it possible that XMRV is ( or related to) an 'escaped,' genetically- engineered laboratory research retrovirus?
  • • A: NO laboratory researchers are not smart enough to engineer new human retroviruses. XMRV has never been detected in ANY other animal in nature except humans.
• [Nancy Allen] Q: Please explain the significance of "xenotropic" in the transmission or infectivity of XMRV ? 
  • • A: Xenotropic viruses are not able to infect cells derived from laboratory mice (or the mice themselves but they can infect cell lines from a number of other species including humans and wild field mice...more proof that this is not a lab engineered or escape variant.
• [Nancy Klimas] Q: Hey guys! miss you! I am keeping things hopping here in Miami, but it isweird being off the board. I noticed VIP Dx has dropped the PCR part of the XMRV testing, in favor of culture. Why?
  • • A: PCR of genomic DNA was found by the WPI and its NCI collaborators to be the least sensitive method of detecting XMRV because of the low copy number of XMRV and the realization by the negative studies that only 1 in 1miiolon resting white blood cells harbors a copy of the XMRV provirus. Thus VIPDx was getting too many false negatives..ie the failure to detect XMRV. The same reason that other companies using sensitive PCR failed to detect XMRV from a spot of blood or 1 ml of blood. Retroviruses multiply only in dividing cells so VIPDx dropped the first PCR in favor of biological amplification of XMRV before PCR detection. Not only are the results more reliable but the test is considerably cheaper. The most important thing is to get the CORRECT answer! 
• Q: Do you feel that XMRV is necessary and sufficient or necessary but not sufficient or not necessary or sufficient to cause chronic illnesses like CFS?
  • • A: Our hypothesis is that chronic XMRV infection creates results in an immune deficiency that is necessary but not necessarily sufficient to result in chronic illnesses like CFS. As with HIV/AIDS copathogens define AIDS, CMV retinitis, kaposi’s sarcoma are AIDS defining co-pathogens. That said XMRV is a type C retrovirus and Type C retroviruses are associated with neurovirulence in a range of host species, including humans, cats, rodents and birds with or without concurrent immunological disease suggesting XMRV can be Necessary and sufficient to cause CFS.
• Q: What are your current % positives in CFS? In controls?
  • • A: In CFS disease satisfying the Canadian consensus criteria, we have isolated virus from more than 300 patients of~400 tested but we have serological evidence in another ~10% of CFS patients fulfilling CCC criteria, from whom we have not isolated virus, clear evidence of infection but the significance of which is not known.
• Q: Is there an indication that the virus is active and being transcribed in CFS? What percentage of positives are being transcribed?
  • • A: The Science paper clearly showed active transcription of XMRV in >75% of the pateitns tested and similar suggestion of latent infection or evidence of infection without isolation of virus in 10-15%.
• Q; With the known % of CFS patients positive for Mycoplasma species (~60% in multiple studies), Chlamydia pneumoniae (~10% in multiple studies), HHV-6 (~30% in some studies) and other infections, is there any concordance with XMRV positivity?
  • • A: We have only done those analyses on the 101 in the original study, HHV6A was 10%, EBV ~14% and nothing else more than 10%. We are working with several groups at Lyme and those numbers may approach 30%-40 of those tested.
• Q: Do you feel that XMRV could act to cause dysfunction of the immune system, allowing opportunistic infections (such as in 4, above), similar to HIV-1 in AIDS?
  • • A: Absolutely that is our working hypothesis
• Q: Do you think that treatment of XMRV might be useful or possibly too late in CFS to restore patients to pre-infection health status?
  • • A; I am extremely optimistic that treatment of XMRV or its immune target(s) will restore CFS patients to at least 85% of the original health status even in the sickest of the sick. I base this optimism solely on the level of health that was achieved in HIV AIDS patients with the advent of highly active antiretroviral therapy (HAART) and XMRV is much less cytopathic than HIV, its effects on the immune system are more subtle. I think the evidence is that some recover with out ever treating the XMRV or maintain relative health for years between “crashes”.
• Q: When you assess signs/symptoms of patients with positive XMRV, are there any that track or don't track well with XMRV positivity compared to CFS patients as a group?
  • • A; I am not a clinician and right now we don’t have sensitive quantitative assays to monitor viral load so we have not actually tracked symptom severity and XMRV positivity. For Science paper we did correlation analyses on every clinical marker of immune status, that we had available to us including RNAase L, NK (number and function, co-infections, cytokines and chemokines and NOTING significantly correlated with a CFS diagnosis (the referees and editors felt these data were not important for the paper as they were all negative data. The only one they published was the R462Q RNaseL variant as this was the original hypothesis under which XMRV was discovered. The ONLY immune marker which correlated with XMRV infection 100% was decreased Interferon alpha.
• Q: In families with more than one CFS (or other chronic illness), is there any indication that XMRV was transmitted to immediate family members? Prof. Garth NicolsonThe Institute for Molecular Medicine
  • • A: YES...lots of them and I have not looked at everyone but my knowledge of the 101 in the Science paper is that more than 2/3 of them have family member who are infected and MOST are ill perhaps with another overlapping chronic illness..for instance FM and CFS or CFS mom and autistic kids and the cancer in some of the families is frightening.
• Q: I read where 75 top researches met on 16 Dec. 2009 to discuss the current research findings about XMRV. I have been looking, but unable to find any information about what they had found. Hope you will include this.William HaysA:I am not aware of that meeting nor am I aware that on December 16th of 2009 there were 75 top researchers working on XMRV! On November 10 the Cleveland Clinic sponsored a meeting where ~15 scientists presented their data including the WPI) on XMRV in cancer and CFS. There were 75 people in attendance and it was an interesting meeting but honestly I would be surprised is there are yet 75 top researchers in the world working on XMRV.Q: How do you see HHV-6 and XMRV interacting? 
  • • A: There are no data to suggest that HHV6 and XMRV directly interact in any way. XMRV is a simple retrovirus so it encodes only structural proteins and not transacting factors like HIV and HTLV. We are testing the hypothesis that like HIV, the transmission of XMRV and the progression of XMRV disease are accompanied by de novo infection by other microbes or by activation of microbes that were present in the host in homeostatic equilibrium before XMRV infection. In recent years, data have accumulated on the interactions of these coinfecting microbes—viruses in particular—with HIV. Coinfecting viruses generate negative and positive signals that suppress or upregulate HIV-1, suggesting that the signals generated by these viruses may largely affect XMRV transmission and pathogenesis as they do with HIV transmission and pathogenesis. 
• Q: The Brits have commented that both M.E., and the U.S. cluster outbreaks of the 1980s, seem to spread like an enterovirus (a polio-like virus), not like a herpes virus.
  • • A: XMRV is a retrovirus not a herpesvirus so I don’t understand the comment. HIV causes leaky gut and disruption of the gut microbiome and an open blood brain barrier allowing pathogens access to areas of the body that these pathogens normally would not have access...the same could happen with XMRV...I guess I cannot answer this because more than 75 of the 101 patients in our study were not involved in cluster outbreaks.Whether that criticism holds, it certainly seems the outbreaks did not follow the pattern set by HIV. While our knowledge of HIV disease is useful in developing hypotheses and studying potential mechanisms of HIV pathogenesis, XMRV is totally unlike HIV and HTLV-1 in that it is a simple retrovirus the first ever identified human exogenous simple retrovirus and the family of viruses most closely related to XMRV causes neurological diseases and cancer very by mechanisms distinct in many aspects from HIV and HTLV. We discussed earlier XMRV may be much more stable in body fluids other than blood and cell free and would completely explain clusters and sporadic transmission as seen in CFS both in the US and UK
• Q: How would teachers in the same school, for example (who were not misbehaving), contract the disease from each other? 
  • • A: See above but what if XMRV is more stable in body fluids such as saliva, urine, vomit such that exchange of body fluids less directly than sexual contact and blood borne direct infection were the mode of transmission.This hypotheses would satisfy the familial and close contact such as a school particularly if a co-pathogen enhanced transmission and progression as was the case of HIV and HSV in Africa.
• Q: What does this mean for young women who wish to become pregnant, but who might have XMRV? 
  • • A: Gammaretroviruses can be transmitted vertically and are transmitted in breast milk. The hormone responsiveness of the virus suggests that lactating moms express more XMRV in breast milk. HTLV 1 was endemic in Japan so simply preventing breast feeding reduced HTLV-1 associated neuroimmune disease and cancer by 40% on one generation. Vertical transmission is more difficult but even if infected, the key is to keep the reservoirs of virus low..Stop the XMRV from being expressed and spread to healthy cells keep the XMRV provirus from integrating into long lived cells ....the variation was so small and the lack of quasi species and high levels of replication suggest that vaccine development will be easier with this retrovirus that with the complex retroviruses, HIV and HTLV. The most important thing to do is identify everyone infected and stop the spread and replication of the virus in every individual prevent disease in those who are well and stop the progression and restore the immune system in those who are ill.
• Q: Given the association of XMRV with patients whose blood was stored at WPI, how many other docs have sent samples that are positive?
  • • A: Contrary to the assumptions and misinformation about the samples in the WPI repository. These samples were NOT solely from the incline village outbreak. All samples were drawn from patients coming to Sierra Internal medicine between 2006 AND 2008. Fewer that 20 of the 101 were from the original outbreak and more than 75 were sporadic cases of CFS from patients who came to Dr. Peterson from 12 states and Canada. The 101 were representative of patients satisfying CCC and Fukuda criteria throughout the US. At least 5 doctors from across the US including Dr Klimas and Dr Cheney had patients in the repository and in the study. The 101 patients were selected at random from the hundreds in the repository. The main consideration was that we had samples that could be cultured (retroviruses cannot multiply unless a cell divides and several samples from a given patient.
• Q: Are your “hit rates” different in the samples sent to you since the Science paper?
  • • A: Not as long as the physicians sending the samples are diagnosing as Dr Peterson does on CCC criteria. In fact the hit rates from overlapping diseases more than we expected now ~35%.
• Q: Given that a number of physicians who send you XMRV samples are also collecting HLA DR data, can we sort the positives and negatives by HLA DR haplotype, i.e., is there evidence of an increased relative risk for carriage of XMRV and presence of illness by HLA DR?
  • • A: We established the original WPI research program to look at the underlying genetics from the exact same population from which we were looking for novel viruses and correlating immune profiles and gene expression patterns. Therefore our collaborators Mary Carrington and Michael Dean of the NCI’s genomic diversity program and the geneticists who identified the HLA and KIR susceptibility and resistance loci in HIV/AIDS have all of these studies well underway...some of these data were presented at last year’s IACFS meeting. At that time we did not know XMRV status of those patients. Those data are currently being analyzed.
• Q: Do we know of any patients with acute acquisition of XMRV? How can you confirm a new case versus re-activation of transcription of intercalated viral material in DNA? What happens to their symptom with acquisition? What happens to their measures of innate immune responses?
  • • A: No. None of these studies can be done yet. We are working on cases of suspected “transfusion acquired” CFS which could establish an acute infection. These studies will require quantitative assays. The requirement that a diagnosis of CFS require an individual to be ill for 6months has precluded anyone from having such data. Hopefully at least one archaic practice will soon be gone ..the worst predictor of HIV disease progression is the viral set point and reservoir viral load. Set the reservoirs as low as possible! That is identify the infection as soon as possible. This is the best chance to prevent disease development and progression.
• Q: Do we know if genes carried by XMRV are being transcribed? If so would genomic data from samples collected by other labs be useful in a pooled analysis? 
  • • A: XMRV is a simple retrovirus and encodes and expresses only the gag pol and env genes. ALL are expressed and proven to make functional proteins. This was clearly demonstrated in our Science paper. Gamma retroviruses do not cart or pick up cellular genes? Hope I answered the question because I don’t understand the second part.
• Q: What regulatory mechanisms are felt to be present that would affect transcription of XMRV genetic material?
  • • A: Expression of XMRV and all simple retroviruses are controlled only by cis acting elements in the upstream untranslated region. In XMRV 3 transcriptional control elements have been identified two gluccocorticoid response elements ( a strong one responding to androgens and estrogens, a weaker one responding to the stress hormone cortisol and an NfKB site) All turn on XMRV.
• Q: If XMRV is actively being transcribed, is there a difference in lab parameters and symptoms compared to those who carry XMRV but are without transcription?
  • • A: We don’t have these data yet but that is a testable hypothesis.
• Q: If XMRV is present but inactive, are there any suggestions as to what could be a trigger for (re)-activation?
  • • A: Estrogens, Androgens, Cortisol (stress) and inflammation.
• Q: Given the problems with antigen presentation seen routinely in so many patients with chronic fatiguing illnesses, we would expect to see some patients with culture (+) XMRV who are antibody negative?
  • • A: Yes! We see lots of those...these data are very interesting and suggest immune therapy including antibody therapy may be most useful in this disease.
• Q: I heard that there may be another study in the U.K. that came to a different conclusion about a possible link between CFS and XMRV. Can you comment on the differing results, and whether the results of the two studies can be reconciled? Thank you. 
  • • A: The negative studies were technically flawed in that their methods were demonstrated NOT to be capable of detecting XMRV. Their patient populations likely did not satisfy CCC criteria and they looked only by PCR on genomic DNA the least sensitive way of detecting XMRV. To date none one has attempted to replicate our study. It is very clear that the prevalence of XMRV in UK is NOT ZERO and that XMRV has been detected in CFS patients in the UK.
• Q: How might the finding of the XMRV virus relate to Lyme Disease?
  • • A: We are seeing XMRV in Chronic Lyme patients sent to us from several physicians. The hypothesis that chronic XMRV infection creates an underlying immune deficiency is consistent with many co-pathogens including Lyme.

Chronic fatigue syndrome causes a host of debilitating symptoms: profound exhaustion, disordered sleep, muscle and joint pain and severe cognitive problems, among others. But what causes the syndrome itself?

Since the first cases in the United States were identified in the 1980s, scientists have been divided over that question. Some have suspected that one or more viral infections are likely to play a central role.

But many other researchers — not to mention relatives, friends, employers, doctors and insurers of the million or more Americans estimated to suffer from the illness — have dismissed it as stress-related, psychosomatic or simply imaginary.

Now recent back-to-back announcements have highlighted both the volatility of the issue and the ambiguity of the science, and have alternately heartened and dismayed patients.

On Dec. 14, an advisory panel suggested that the Food and Drug Administration ban blood donations by people with a history of C.F.S., as the illness is often called. The goal was to prevent the possible spread of viruses that two high-profile studies had linked to the condition.

But then, on Dec. 20, the journal Retrovirology published four papers suggesting that key findings in those studies could have resulted from laboratory contamination.

The F.D.A. is not required to accept the opinion of its advisory panel. Yet patients still hailed the recommendation as a sign that their illness was being taken seriously.

“When an F.D.A. panel suggests that patients with C.F.S. not donate blood, that’s going to impact the way doctors think about it,” said Mary Schweitzer, a former history professor at Villanova, who has frequently written about living with the illness. Dr. Schweitzer said she has been unable to work for 16 years because of the syndrome, which was diagnosed after she suffered from a series of flulike illnesses.

The studies that concerned the F.D.A. had reported that people with the syndrome, which is also called myalgic encephalomyelitis or myalgic encephalopathy in Europe, showed higher rates of infection with the virus XMRV or others from the same category, known as MLV-related viruses. (These viruses are all relatives of mouse leukemia viruses, some of which can infect species other than mice; their role in human disease, if any, remains poorly understood.)

But several other research teams in the last year have found no connection between chronic fatigue syndrome and these viruses, although none tried to replicate the exact methods used by researchers who reported an association.

The new papers in Retrovirology reported that contamination of tissue samples or other laboratory items with mouse DNA or viral genetic material could lead to false positive results for XMRV, and by extension other MLV-related viruses, specifically when using polymerase chain reaction technology. The technique rapidly produces millions of copies of genetic segments, so even minute traces of genetic contamination can skew results.

“Our conclusion is quite simple: XMRV is not the cause of chronic fatigue syndrome,” said the senior author of one of the studies, Greg Towers, a professor of virology at University College London, in a statement released by Wellcome Trust Sanger Institute, the British research center that co-sponsored it.

Other scientists and advocates for patients have sharply criticized such certainty as unwarranted, noting that the Retrovirology papers themselves expressed their findings in more cautious terms. The critics agree that contamination can be a serious issue when using polymerase chain reaction technology. But the new papers, said Eric Gordon, a doctor in Santa Rosa, Calif., who treats many patients with the illness, do not evaluate other strategies besides P.C.R., as the technique is known, for detecting the MLV-related viruses, like testing for an immune response and culturing the viruses in cell lines.

“The articles make the point that P.C.R. doesn’t work that well for these viruses, and then they act like that disproves the whole idea,” said Dr. Gordon.

XMRV was first identified in 2006 and has been detected in prostate cancer patients in some studies. It was linked to chronic fatigue syndrome in October 2009 in a paper in the journal Science by researchers from the Whittemore Peterson Institute for Neuro-Immune Disease at the University of Nevada, Reno, the National Cancer Institute and the Cleveland Clinic.

The researchers relied on P.C.R. technology to show that about two-thirds of patients but less than 4 percent of control subjects harbored XMRV. Using other technologies, however, they also documented an antibody response in some chronic fatigue syndrome patients, and reported that XMRV in human blood could infect other human cell lines.

In a statement responding to the new papers in Retrovirology, Judy A. Mikovits, director of research at Whittemore Peterson and the senior author of the Science study, said her team took extensive steps to rule out P.C.R. contamination and also focused on other approaches to finding XMRV. “Nothing that has been published to date refutes our data,” she said.

Even some specialists stumbled over the meaning of the new findings. Vincent Racaniello, a professor of microbiology at Columbia not involved in the research, apologized on his Virology Blog for having stated that it was likely to spell “the beginning of the end” for the proposed connection between the viruses and chronic fatigue syndrome.

After reviewing the issue more thoroughly, he wrote, he realized that the new studies “show that identification of XMRV can be fraught with contamination problems, but they do not imply that previously published studies are compromised.” He added, “If I had difficulties interpreting these papers, how would nonscientists fare?”

Federal agencies have come down on different sides of the issue. In a paper published in The Proceedings of the National Academy of Sciences in August, researchers from the National Institutes of Health and the F.D.A. found a link between the fatigue syndrome and MLV-related viruses (although not specifically XMRV). In contrast, a study from the Centers for Disease Control and Prevention was among those not reporting a link.

Federal health officials have organized two research efforts to resolve the inconsistencies, determine whether XMRV and MLV-related viruses are possible human pathogens, and identify reliable ways to detect them. Patients hope the increased attention will quickly lead to research on treatments, including clinical trials of H.I.V. drugs, some of which have been shown in lab studies to inhibit the replication of XMRV.

The unsettled situation has created a quandary for patients with chronic fatigue syndrome and the doctors who treat them. Some patients are seeking to be treated with H.I.V. drugs, which doctors can legally prescribe even though the F.D.A. has not approved them for that purpose.

Many doctors and researchers say it is too early to prescribe the drugs for chronic fatigue because of possible side effects, like bone marrow suppression, gastrointestinal problems and liver or kidney dysfunction, among others. But Michael Allen, a writer and a former psychologist in San Francisco who has been disabled for more than 15 years, said he wouldn’t hesitate to try the medications if he found out he was positive for an MLV-related virus.

“It feels patronizing when the medical establishment says the side effects are too risky and we should keep waiting,” he said. “What that says to me is they have no idea whatsoever how sick people like me have been with this disease.”

In a blow to patients with chronic fatigue syndrome, two new studies published on Tuesday raised serious doubts about earlier reports that the disabling disease is linked to infection with XMRV, a poorly understood retrovirus.

The new papers were posted online in the journal Science, which in October 2009 published the initial research linking XMRV to chronic fatigue syndrome. In an “editorial expression of concern” accompanying the two new studies, Bruce Alberts, editor in chief of the journal, declared that the earlier finding “is now seriously in question” and was most likely due to laboratory contamination.

Based on those earlier findings, some people with chronic fatigue syndrome tried to obtain access to antiretroviral drugs used to treat H.I.V., which had been shown in laboratory studies to inhibit the replication of XMRV.

But in one of the two new studies, researchers found no trace of XMRV or related viruses in the blood of 43 patients who had previously tested positive for XMRV. In the second study, scientists reported evidence that XMRV was likely a recombination of two mouse leukemia viruses created accidentally in laboratory experiments.

The new studies are the latest in a series of disappointments for people struggling with chronic fatigue syndrome. Other researchers have been unable to duplicate the original findings implicating XMRV, although none of their studies fully replicated the methods of the original research from the Cleveland Clinic, the National Cancer Institute and the Whittemore Peterson Institute for Neuro-Immune Disease in Reno, Nev

Dr. Vincent Racaniello, a microbiology professor at Columbia University, said in an interview that it now appeared unlikely that XMRV infection is a cause of chronic fatigue syndrome. But it also would be wrong to conclude that chronic fatigue syndrome is not an infectious disease, he added.

“These patients have a lot of signs of hyper-immune activation, with their immune systems firing almost constantly,” he said

[Dr. Jay A. Levy (born 1938)], a professor of medicine at the University of California, San Francisco, and the senior author of one of the new studies, said he nonetheless believed that many or most people with chronic fatigue syndrome are suffering from a disease initiated by one or more viruses.

Many of the disease’s symptoms are likely caused by the immune systems’ response to an infection, rather than to the pathogens themselves, he said.

“The immune system pours out its toxins to stop this agent, and then the immune system doesn’t calm down,” he said, adding that environmental toxins could also play a role in the illness.

Last week, the editors of Science asked the authors of the original research if they would retract their paper in view of the new findings about to be published. The senior author, Dr. Judy Mikovits, research director at the Whittemore Peterson Institute, responded that such a step was “premature” and that she knew of other investigators planning to publish research backing the original findings.

Some patients reacted angrily to the news that Science had asked for a retraction. “The patient community is shocked,” Rivka Solomon, a largely homebound Boston-area writer with chronic fatigue syndrome, said in an e-mail. “Most of us feel that the scientific inquiry necessary to bring this to conclusion has not yet been played out.”

Ms. Solomon recently organized small demonstrations in Washington, D.C., and San Francisco to focus attention on the small amount of financing the government has allocated to the disease in recent years. If the XMRV association does not pan out, she wrote, patients like her “worry that we will once again be abandoned.”

The government is supporting additional studies to determine whether XMRV plays any role in chronic fatigue syndrome at all. The retrovirus had also been linked to prostate cancer, an association also challenged by the research published on Tuesday.

Judy Mikovits has had a rough few weeks. On 22 September, Science published online a nine-lab study widely seen as the final blow to the theory,  championed by Mikovits and colleagues in an October 2009 Sciencepaper, that a recently detected mouse retrovirus might play a causal role in chronic fatigue syndrome (CFS). A letter in the same issue of Science from one of the contributing labs to the 2009 report revealed that a contamination had marred its contribution—PCR detection and sequencing of the mouse virus, dubbed XMRV. Mikovits and colleagues defended the validity of the rest of the study, known as Lombardi et al., which detected the virus by several other methods, so Science  issued a rare partial retraction of the original paper. 

Then on 29 September, Mikovits was fired from her job as research director of the Whittemore Peterson Institute for Neuro-Immune Disease (WPI), a private organization in Reno, Nevada, devoted to CFS research and treatment. Both Mikovits and WPI's CEO, Annette Whittemore, say the firing was not related to the XMRV theory's demise. 

The very next day, a graduate student who writes a snarky blog that has been highly critical of Mikovits and the XMRV theory raised questions about whether a figure in Lombardi et al. had been misrepresented. Science Executive Editor Monica Bradford said in a statement that the journal is investigating the allegation. "As is our policy in cases of alleged figure manipulation, we follow up with the research authors as soon as our own review of the allegation is complete," said Bradford. "Science takes all such matters seriously and seeks to respond thoroughly and efficiently."

The furor revolves around an image—the bottom half of Figure 2C in Lombardi et al.—that shows XMRV proteins in CFS patients but not healthy controls. In her blog known as ERV (endogenous retroviruses), Abbie Smith on 30 September noted the striking similarities between Figure 2C and a slide  Mikovits presented at a CFS meeting in Ottawa, Canada, on 23 September. Smith, who is working on her doctoral dissertation at the University of Oklahoma, Oklahoma City, and studies HIV, wrote that an anonymous tipster had pointed out to her that the two images looked identical but had different patient numbers and experimental conditions. Smith questioned whether this was a simple mistake or an attempt to recycle old data to make a new argument. 

The Ottawa slide supported Mikovits's contention that even if XMRV could not be detected in CFS patients, other gammaretroviruses still lurked in their chromosomes. Mikovits described how she had treated cells from two CFS patients with a chemical, 5-azacytidine, that takes methyl groups off DNA. This procedure prods cells that harbor latent versions of retroviruses to produce them, and the image on the slide showed the resultant proteins in what's known as a Western blot gel. In Lombardi et al. what appears to be the same image shows "XMRV proteins" and makes no mention of 5-azacytidine use. 

Mikovits's collaborator, [Dr. Francis William Ruscetti (born 1943)] of the National Cancer Institute (NCI) in Frederick, Maryland, who ran all of the Western blots, confirms that the Ottawa slide uses the same image that appears in Lombardi et al. Ruscetti and Mikovits, in a joint e-mail to Science for this article, said many patients and their doctor, Daniel Peterson (who since has had a falling out with WPI), knew the original coded numbers, so the researchers changed them for the Science publication to "protect the patient privacy." Ruscetti says it was a mistake for Mikovits to have used the original patient codes in Ottawa. "We were under so much pressure, we missed it," says Ruscetti. 

As far as the use of 5-azacytidine, Ruscetti and Mikovits stressed in their e-mail that "there was no attempt in the original paper to hide anything." They say for the purposes of Lombardi et al., the use of 5-azacytidine was not germane: They were simply trying to demonstrate that CFS patients had viral proteins not seen in controls. By the time of the Ottawa meeting, they say they realized that this experiment did not in fact show XMRV but proteins from a broader family of gammaretroviruses.

After Lombardi et al. appeared, several laboratories had reported that they could not detect XMRV in CFS patients. But [Dr. Francis William Ruscetti (born 1943)] and Mikovits note that most of these studies relied on polymerase chain reaction, which used DNA sequences of XMRV to fish out pieces of the virus from blood samples. Those tests, they point out, would have missed other gammaretroviruses with different sequences. The Western blot assay they used in Lombardi  et al. just so happened to cast a wider net that uncovered proteins from any member of the gammaretrovirus family. In addition, they say, the use of 5-azacytidine made clear that these infections would be missed in routine assays, as these viruses often exist in a latent state. 

On the ERV blog, Smith and others also argued that the study lacked proper controls: the healthy controls in this experiment did not have 5-azacytidine added to their samples. "My [principal investigator] would say 'Why did you run this gel?' if I handed it to him," wrote Smith. 

Vinay Pathak, a retrovirologist at NCI who earlier damaged the XMRV/CFS theory with a study published in Science that  documented how the virus was accidentally created in laboratory experiments, says he is "bewildered" by Ruscetti's and Mikovits's explanations about Figure 2C. "If [5-azacytidine] was used in the original experiment, it's an egregious error to leave it out of the Science paper," says Pathak. "It makes a difference how I would interpret the results." 

Jonathan Stoye, a retrovirologist who once supported the XMRV/CFS hypothesis but subsequently changed his mind after his own studies failed to replicate the finding, says its time to retract Lombardi et al. in its entirety. "I think there's a point where Science has to say, there is no substance to this paper," says Stoye. "It was published with a message, and that message is gone." 

Neither Lombardi et al. nor the questions about the slides were mentioned in Whittemore's 30 September letter formally terminating Mikovits's contract, sent the day after the two had a heated phone conversation. In that letter Whittemore charged her high-profile researcher with being "insubordinate and insolent." Mikovits was immediately locked out of her lab. 

Three letters between Whittemore and Mikovits say the firing hinged on Mikovits's failure to pass on a cell line that was sent to Vincent Lombardi, the first author of the October 2009 Science paper who runs UNEVX (formerly known as VIPDx), a diagnostic laboratory owned by WPI. Until recently, the lab sold a test for XMRV and related viruses.

In a 1 October written response to Whittemore, Mikovits contended that it was "completely appropriate" for her, as research director, not to give Lombardi the cell line. The cell line was not related to studies of the gammaretroviruses, but Lombardi wanted to use it for experiments connected to a grant Mikovits had secured from the U.S. National Institutes of Health to study possible causes of CFS. Mikovits contended that Lombardi "was unwilling to take my direction" and should not be undertaking a new project "while neglecting his other duties." She also questioned his ability to carry out that experiment. 

Annette Whittemore issued a statement to Science in which she strongly defended Lombardi's performance. "Dr. Lombardi is a valued and important part of our team, and conducts his research work accordingly," wrote Whittemore. "While personnel matters are generally confidential, the statements made by Judy Mikovits are wrong, without merit and those of a disgruntled former employee." 

In an interview with Science, Mikovits contended that her firing was also linked to a longstanding battle about WPI's decision to sell, through VIPDx/UNEVX, a test for human gammaretroviruses. The lab began offering the tests, which cost around $500, shortly after Lombardi et al. reported a link between XMRV and CFS. Some patients who tested positive went on to take antiretroviral drugs. "I said, 'No, no, no, no,' " says Mikovits of the test. "I've asked them for the better part of 2 years to show me that what we got in Lombardi et al. is the same thing they're selling to patients." 

The issue came to a head with the recent publication by Science of the nine-lab study. The so-called Blood Working Group, which included the labs run by Mikovits and [Dr. Francis William Ruscetti (born 1943)], failed to reliably find XMRV or other gammaretroviruses in blinded samples from people who previously had tested positive for these viruses. Both Mikovits and Ruscetti co-authored the paper, which invalidated their own assays for XMRV. WPI says UNEVX has stopped offering the diagnostic tests, but did not give details about the timing or reasons.

Whittemore, who refers to CFS as myalgic encephalomyelitis (ME)—a common name for CFS in Europe—stressed that the institute remains devoted to studying human gammaretroviruses in "ME and related diseases" and that no one there "would ever put self interest ahead of research or finding the causes of ME." 

Mikovits, who says she currently does not even have access to her laboratory notebooks, is looking for another institution to continue her work.

When scientists reported in 2009 that a little-known mouse retrovirus was present in a large number of people with chronic fatigue syndrome, suggesting a possible cause of the condition, the news made international headlines. For patients desperate for answers, many of them severely disabled for years, the finding from an obscure research center, the Whittemore Peterson Institute for Neuro-Immune Disease in Reno, Nev., seemed a godsend.

“I remember reading it and going, ‘Bingo, this is it!’ ” said Heidi Bauer, 42, a mother of triplets in Huntington, Md., who has had chronic fatigue syndrome since her 20s. “I thought it was going to mean treatment, that I was going to be able to play with my kids and be the kind of mom I wanted to be.”

Patients showered praise on the lead researcher, Dr. Judy Mikovits, a former scientist at the National Cancer Institute. They sent donations large and small to the institute, founded by Harvey and Annette Whittemore, a wealthy and politically well-connected Nevada couple seeking to help their daughter, who had the illness.

In hopes of treating their condition, some patients even began taking antiretroviral drugs used to treat H.I.V., a retrovirus related to the murine leukemia viruses suddenly suspected of involvement in chronic fatigue syndrome.

More recently, however, the hopes of these patients have suffered an extraordinary battering. In a scientific reversal as dramatic and strange as any in recent memory, the finding has been officially discredited; a string of subsequent studies failed to confirm it, and most scientists have attributed the initial results to laboratory contamination. In late December, the original paper, published in the journal Science, and one other study that appeared to support it were retracted within days of each other.

As the published evidence for the hypothesis fell apart, a legal melodrama erupted, dismaying and demoralizing patients and many members of the scientific community. Dr. Mikovits was even briefly jailed in California on charges of theft made by the institute.

“I’m stunned that it’s come to this point,” said Fred Friedberg, a professor at Stony Brook University Medical Center and president of the International Association for C.F.S./M.E., a scientific organization. “This is a really sad unraveling of something that was perhaps going to generate a whole new direction in this illness.”

Despite the controversy, Dr. Mikovits is now supervising some lab work as part of a large government-sponsored study being spearheaded by [Dr. Walter Ian Lipkin (born 1952)], a leading Columbia University virologist. The study was established before the two retractions to examine the possible link between chronic fatigue syndrome and mouse retroviruses. Dr. Mikovits still hopes to replicate her original results, and many patients continue to believe fervently in her hypothesis; study results are expected early this year.

She did not respond to requests for comment.

An estimated one million people in the United States suffer from chronic fatigue syndrome, which is characterized by profound exhaustion, a prolonged loss of energy following minimal exertion, swollen lymph nodes, sore throat, cognitive dysfunction and other symptoms. Experts now generally believe that one or more infectious agents, or perhaps exposure to toxins, set off a persistent, hyperactive immune response — the likely cause of many of the symptoms.

Although the Centers for Disease Control and Prevention first investigated the illness in the mid-1980s, the agency has not been able to find a cause, identify any biomarkers or diagnostic tests, or develop effective treatments. Patients have long accused the mainstream medical and scientific community of neglect and abandonment. Many say that the C.D.C. has largely treated their disease as a psychological or stress-related condition.

A 2010 paper from the agency, for example, galled patients with the conclusion that they suffer disproportionately from “paranoid, schizoid, avoidant, obsessive-compulsive and depressive personality disorders.”

Dr. Mikovits’s research, done with collaborators from such prestigious organizations as the National Cancer Institute and the Cleveland Clinic, seemed to vindicate the concerns of many with the condition. The scientists said they had found that 67 percent of patients sampled were infected with a mouse virus called XMRV, compared with 4 percent of the controls.

“If for years you’ve been told that your illness is all in your head and then you’re being told, ‘Look, we found something concrete and very substantial,’ then of course there will be rallying behind that,” said Rivka Solomon, 49, a Massachusetts playwright who has been largely homebound with the syndrome for more than 20 years.

The publication of Dr. Mikovits’s work brought immediate attention, much of it unflattering. Other scientists soon published studies challenging the findings, and Science issued first a statement of concern and then a partial retraction of the original study.

Even as her work was publicly debated, the blunt and feisty Dr. Mikovits raised eyebrows among other scientists for stating at conferences that murine leukemia viruses could be related to autism. Perhaps more disconcerting, a commercial lab associated with the Whittemore Peterson Institute began marketing screening tests for XMRV, the hypothesized cause of chronic fatigue syndrome, costing hundreds of dollars. The business enraged many patients once they realized the results might be meaningless.

Amid mounting concerns, Dr. Mikovits left her position as research director at the institute in a dispute over management practices and control over research materials. The institute sued her, accusing her of stealing notebooks and other proprietary items. Dr. Mikovits was arrested in Southern California, where she lives, and jailed for several days, charged with being a fugitive from justice.

After her split with the institute, Dr. Mikovits denied having the missing laboratory materials. But a lab employee, Max Pfost, said in an affidavit that he took items at her request, stashing notebooks in his mother’s garage in Sparks, Nev., before turning them over to Dr. Mikovits.

At one point, “Mikovits informed me that she was hiding out on a boat to avoid being served with papers from W.P.I.,” Mr. Pfost said in the affidavit. Some lab items have since been returned.

In December, a judge ruled against her in the civil case. The criminal case is pending; another hearing in the civil case is in late February. But in late January, the Whittemores were themselves accused of embezzling millions of dollars in a lawsuit filed by partners in Mr. Whittemore’s real estate business.

The Whittemores, who have countersued, maintain their innocence of the embezzling charges, and Annette Whittemore stated in an e-mail that institute research continues.

The events of the past couple of years, though disheartening to chronic fatigue syndrome patients, may have a silver lining: Research into the disease, much of it privately financed, is ratcheting up.

A new research and treatment center has been created at Mount Sinai Hospital in New York. The Hutchins Family Foundation is investing $10 million in the Chronic Fatigue Initiative, an effort to find causes and treatments that has recruited top researchers from Columbia, Harvard, Duke and other institutions.

“The disease had languished in the background at N.I.H. and C.D.C., and other scientists had not been paying much attention to it,” said John Coffin, a professor of molecular biology at Tufts University. “This has brought it back into attention.”

Dr. Coffin, who at first supported the mouse retrovirus theory but later disputed it, noted that the illness “does seem to have characteristics that would suggest infectious origins” and that other retroviruses could be involved.

Despite the personal and professional setbacks for Dr. Mikovits, many patients, like Ms. Solomon, continue to believe that a retrovirus is causing their illness.

“But even if the retroviral research does not pan out, her work, and the publicity it has brought to our illness, has forever changed the landscape,” said Ms. Solomon.

Judy Mikovits, who faced a series of professional and legal woes after publishing a study in Science more than 2 years ago that linked chronic fatigue syndrome (CFS) to a mouse retrovirus, has had a pending criminal case against her dropped. 

Last November, the district attorney in Washoe County, Nevada, filed a criminal complaint against Mikovits that charged the virologist with illegally taking computer data and related property from her former employer, the Whittemore Peterson Institute for Neuro-Immune Disease (WPI) in Reno, Nevada. In a separate civil court that month, WPI filed suit against Mikovits over her alleged possession of similar material, which included the laboratory notebooks she compiled while doing the CFS research. 

On 11 June, the district attorney's office for Washoe County filed a petition to dismiss the criminal charges against Mikovits without prejudice (which means they can file a related complaint in the future), a clerk to the Justice Court of Reno told ScienceInsider. 

Mikovits, who was briefly jailed on the charges, is still defending herself in the civil case, which has taken several bizarre twists, including a judge who had ruled against Mikovits recusing himself. The judge removed himself from the case because he received campaign donations from WPI co-founder Harvey Whittemore, who himself has been criminally charged with making illegal campaign contributions to a federal official. (He pled not guilty on 7 June). 

Assistant District Attorney John Helzer, who filed the dismissal, says Whittemore's legal troubles factored into his decision. "There's a lot going on with the federal government and different levels that wasn't occurring when we first became involved with prosecuting this case," says Helzer. "And we have witness issues that have arisen." 

Science last December retracted the 8 October 2009 CFS paper by Mikovits and co-authors, an action she opposed. 

Mikovits told ScienceInsider that the only work she has been able to find has been collaborating on a large study funded by the National Institutes of Health that should be the final word on the otherwise dismissed theory that CFS is linked to a mouse retrovirus, XMRV, or its relatives. "Everyone who wanted to work with me 

was deterred by the threat of litigation," Mikovits wrote in an e-mail. 

The results of the large study, led by Ian Lipkin of Columbia University, are expected to be revealed in the next few weeks

Nevada authorities have dropped criminal charges against a researcher who gained wide attention for scientific reports linking chronic fatigue syndrome to an obscure mouse retrovirus.

The researcher, Judy A. Mikovits, had been charged with stealing proprietary laboratory materials from her former employer, the Whittemore Peterson Institute for Neuro-Immune Disease in Reno, Nev. She still faces related civil claims that the institute filed after her departure.

In October 2009, the journal Science published a study, with Dr. Mikovits as senior author, indicating that a disproportionate percentage of patients with the syndrome were infected with a mouse retrovirus, XMRV. But subsequent studies failed to confirm the initial findings. Other scientists argued that laboratory contamination was the cause of the apparent association. 

As the hypothesis fell apart, so did the relationship between Dr. Mikovits and the institute. Dr. Mikovits left in September. In November, she was arrested and briefly jailed as a result of criminal charges filed by prosecutors in Washoe County, Nev., after the institute’s allegations of theft.

In December, Science retracted her original paper. She is still involved in a large chronic fatigue syndrome study, whose findings are due this summer.

Harvey and Annette Whittemore, who founded the institute, have themselves been sued by former partners, who accused them of embezzling millions of dollars from a real estate business. Mr. Whittemore was indicted last week by federal prosecutors on charges of making illegal campaign contributions. He pleaded not guilty. 

In March, the judge in the civil case against Dr. Mikovits disclosed that he had received campaign donations from Mr. Whittemore and recused himself.

Innovative Cellular and Molecular Biologist with over 20 years of scientific expertise as a PhD in life sciences, including planning, directing and implementing programs in HIV, Cancer, Epigenetics, Neuro-immune disease with a focus on development of novel drug and diagnostic technologies. Demonstrated leadership in introducing and establishing new programs and technologies, efficiently organizing and standardizing processes, and effectively managing multiple projects and personnel. Solution-oriented team-player with strong supervisory, project management, and problem solving skills. Ability to build solid multicultural teams within start-up and changing environments and demonstrated ability to foster strong relationships with strategic partners, collaborators, cross-functional teams, and scientific advisors.

PROFESSIONAL EXPERIENCE

P R O F E S S I O N A L  E X P E R I E N C E  

• MAR CONSULTING INC.   ( 2013 - present )

  • Founder/Consultant  

  • Mission Statement 

  • MAR Consulting Inc., led by [Dr. Francis William Ruscetti (born 1943)] and Dr. Judy Mikovits, seeks to understand complex and innovative biological issues to yield unbiased integrated, cutting-edge information for  patients and physicians impacted by some of the most challenging chronic diseases. Utilizing their combined 75 years experience in tumor biology, immunobiology of retroviral-associated inflammatory diseases, cancer, stem cell biology, hematopoiesis, and drug development, MAR focuses on research projects, consulting (to patients doctors, academia, and industry) and lecturing without the restrictive authority of vested interest groups, following Thomas Jefferson's dictum: "Here we are not afraid to follow the truth wherever it may lead, nor to tolerate error so long as freedom is left free to combat it."

• YORKBRIDGE CAPITAL     (  2012 - present )

  • Advisor  

  • York Bridge Capital is a Toronto-based private equity firm focused on early stage Canadian technology companies. My role is in management advisory services for the medical devices and diagnostics sector.  

• GENYOUS BIOMED   (  2006 - 2012 )

  • Senior Scientist/Consultant  

  • As a senior scientist I developed multivalent drugs for inflammation, immune modulation and virology targets. Genyous’  Multifunctional Multi-targeted (MFMT™) drug development platform is based on systems biology considerations that recognize the complex nature of chronic diseases and addresses the crosstalk between cells, their microenvironment (stroma) and the immune response system. Genyous’ drug candidates are formulated with well-tolerated doses of actives derived from natural products that have a long history of safe consumption and which are orally bioavailable. MFMT™ drug design also minimizes disruption of homeostasis, thus reducing risk of toxicity and development of drug resistance. Genyous' multivalent drugs are designed to address the heterogeneous nature of chronic diseases by possessing multiple therapeutic functions and acting on multiple biological targets. As a consultant, I interacted with academic collaborators world-wide to establish milestones for MFMT research in prostate cancer, autoimmune and neuroimmune disease.  

• Whittemore Peterson Institute For Neuroimmune Disease (WPI)   (   2006 - 2011   )

  • Research Director, (11/2006- 10/2011)

  • I joined the Whittemore Peterson Institute for Neuro-immune Disease (WPI) in November of 2006 as the WPI’s first Research Director. Responsible for establishing a translational research program aimed at identifying biomarkers and underlying causes of Chronic Fatigue Syndrome (CFS) and other debilitating neuro-immune diseases with overlapping symptoms such as Fibromyalgia (FM), Chronic Lyme Disease, Atypical Multiple Sclerosis and Autism Spectrum Disorder (ASD). As Research Director, I was responsible for planning, establishing and directing the Institute’s scientific research program, including the selection, training and supervision of staff, writing, obtaining and managing grants and collaborating with other scientific organizations. The WPI under my direction grew from a small foundation to an internationally recognized center for the study of neuro-immune diseases in which I obtained investigator initiated grant money from the NIH and Department of Defense and brought international attention to ME/CFS as a physiological disease. Proven ability to recognize and recruit important collaborators and partners developing strong relationships within multiple scientific disciplines.      

• EPIGENX BIOSCIENCES, SANTA BARBARA, CA      (  2005 - 2006 )   [ EpiGenX Pharmaceuticals ]

  • A holding company for technologies developed in EpiGenX Biosciences

  • Chief Scientific Officer,

  • This company was formed to market intellectual properties and technologies developed in EpiGenX pharmaceuticals in order to get treatments to patients faster. As Chief Scientific officer, my job was to identify suitable business partners for product commercialization. We achieved this goal in less than two years and succeeded by having the lead compound acquired by a major pharmaceutical company and into clinical trials and our diagnostic platform licensed to a platform company.  

• EPIGENX PHARMACEUTICALS, SANTA BARBARA, CA     ( 2001 - 2004 ) [ EpiGenX Pharmaceuticals ]

  • A startup biotech company whose mission was to develop and commercialize cancer therapeutics and Diagnostics targeting aberrant DNA methylation in Cancer.

  • Director of Cancer Biology,  

  • Established, directed and managed a team charged with discovering and developing small molecule therapeutics targeting aberrant DNA methylation. In order to achieve this mission we needed first to develop multiplexed, higher throughput assays for determining methylation status of cellular genes.  

• BIOSOURCE INTERNATIONAL, Camarillo, CA (Now part of LIFE TECHNOLOGIES)     2002 - 2004

  • A biotechnology company, which manufactures, markets, and distributes assays and biological reagents for drug discovery, functional proteomics, and biomedical research.

  • Group Leader, Luminex Platform Research & Development. 

  • Organized and directed a team for new product development on the Luminex platform technology for multiplex proteomics assays. Coordinated development and manufacturing of over 50 distinct Luminex assays including the first 30plex assay for human cytokines and chemokines. Responsibilities also included evaluation of external multiplex proteomic technologies and applications for corporate partnership, OEM opportunity and licensing, which led to a contract to commercialize a technology for development of a fully automated platform technology for both multiplex gene expression and proteomic assays on a single platform.  

• LABORATORY OF ANTIVIRAL DRUG MECHANISMS    (  1999 - 2001  )

  • Screening Technologies Branch, a division of the Developmental Therapeutics Branch of the National Cancer Institute, Frederick MD  

  • Lab Director,  

  • Established and managed a new laboratory to develop drugs targeting AIDS-associated malignancies, the first of which was Kaposi’s Sarcoma, which had been recently associated with a new herpes virus. I hired a multidisciplinary team with expertise in high throughput screening, retroviruses, herpes viruses and medicinal chemists. This laboratory became internationally recognized as the first of its kind in cancer drug development developing multi-functional drugs to target pathogens as well as inflammation to fight cancer.  

• NATIONAL CANCER INSTITUTE, LAB OF LEUKOCYTE BIOLOGY    ( 1994 - 1998   )

  • Staff Scientist  

  • Established and managed an independent research laboratory involved mechanisms by which retroviruses disrupt the  delicate balance of the immune system to contribute to disease. Pioneered the field of epigenetic dysregulation of cellular genes by human retroviruses  

• NATIONAL CANCER INSTITUTE, LAB OF GENOMIC DIVERSITY    (  1992 - 1994   )

  • Post Doctoral Fellow, Molecular Virology

  • Constructed and characterized the first infectious molecular clone of HTLV1. Investigated the role of defective provirus in HTLV-1 associated myelopathy (HAM/TSP)

• NATIONAL CANCER INSTITUTE, LAB OF LEUKOCYTE BIOLOGY                             1983 - 1991  

  • Research Technician II-III,Research Associate, conducted biological and molecular experimentation in HIV AIDs and cancer as a part of the Biological Response Modifiers Program the NCI’s first translational research program.  

• NATIONAL CANCER INSTITUTE, FERMENTATION CHEMISTRY PROGRAM             1980 - 1982  

  • Research Technician I, purification of Interferon alpha, Interleukin 2, and numerous chemotherapeutics for human clinical trials.  

EDUCATION  

• Postdoctoral Scholar: Molecular Virology

• Laboratory of Genomic Diversity, National Cancer Institute with [Dr. David Daniel Derse (born 1949)]

• Ph.D., Biochemistry and Molecular Biology, George Washington University, Washington, DC Doctoral Thesis: HIV Latency and mechanisms of Immune activation In Monocytes

• B.A., Biology with specialization in Biochemistry. University of Virginia, Charlottesville, VA  

PROFESSIONAL SOCIETIES AND AWARDS

• 2012 HHS Special recognition award

• AACR

• DNA Methylation Society

• AAI

• 1991 George Washington Graduate student of the year    

PUBLICATIONS (SELECTED OF 51)

• Farrar, W. L., and Ortaldo, J. R.: Analysis of effector mechanisms against HTLV-I and HTLV-III/LAV infected lymphoid cells. J.Immunol. 136: 3619-3624, 1986.  
• [Dr. Francis William Ruscetti (born 1943)]. and Mikovits, J. A.: Differential regulation of the two IL 2 binding proteins. In Cruise, J. and Lewis, R. (Eds.): Year in Immunology, 1988. Basel, Karger, 1989, pp. 38-45.  
• Mikovits, J. A., Raziuddin, Gonda, M., Ruta, M., Lohrey, N., Kung, H-F. and Ruscetti, F.: Negative regulation of HIV replication in monocytes: Distinctions between restricted and latent expression in THP-1 cells. J. Exp. Med. 171: 1705-1720, 1990.  
• Raziuddin, Mikovits, J., Clavert, I., Ghosh, S., Kung, H-F. and Ruscetti, F.: Negative regulation of HIV-1 expression in monocytes: Role of the 65+50 kD NF kB heterotetramer. Proc. Natl. Acad. Sci. 88: 9426-9430, 1991.
• Mikovits, J. A., Lohrey, N. C., Schuloff, R., Courtless, J. and Ruscetti, F. W.: Immune activation of HIV expression from latently infected monocytes from asymptomatic seropositive patients. J. Clin. Invest. 90: 1486-1491, 1992.  
• Hoffman, P. M., Dhib-Jalbut, S., Mikovits, J. A., Robbins, D. S., Wolf, A. L., Bergey, G. K., Lohrey,N., Weislow, O. S. and Ruscetti, F. W.: HTLV-1 infection of monocytes and microglial cells in primary human cultures. Proc. Natl. Acad. Sci. 89: 11784-11788, 1992.
• West, M. Mikovits, J., Princler, G., Liu, Y-L., Ruscetti, F., Kung, H-F. and Raziuddin.: Characterization and purification of a novel transcriptional repressor from Hela cell nuclear extracts recognizing the negative regulatory element of HIV-1 long terminal repeat. J. Biol. Chem. 267: 24948-24952, 1993.  
• Li, C-C., Ruscetti, F., Rice, N., Chen, E., Mikovits, J., Yang, N-S. and Longo, D. L.: Differential expression of Rel family members in HTLV-1 infected cells: Transcriptional activation of c-rel by tax protein. J. Virol. 4205-4213, 1993. 
• Mikovits, J. A., Meyers, A. M., Ortaldo, J. R. and Ruscetti, F. W.: IL-4 and IL-13 have overlapping but distinct effects on HIV production in monocytes. J. Leukocyte Biol. 56: 340-346, 1994.  
• Mayers, D. L., Mikovits, J. A., Joshi, B., Hewlett, I. K., Pankaskie, M. C., Estrada, H. S., Wolfe, A. D., Garcia, G. E., Buyke, D. S., Gordon, R. K., Lane, J. R. and Chiang, P. K.: Novel anti-HIV-1 activities of 3-deaza adenosine analogs: Increased potency against AZT-resistant HIV-1 strains. Proc. Natl. Acad. Sci. 92: 215-219, 1995.  
• [Dr. David Daniel Derse (born 1949)], Mikovits, J., Polianova, M., Felber, B. K., and Ruscetti, F. W.: Virions released from cells transfected with a molecular clone of HTLV-1 give rise to primary and secondary infections of T-cells. J. Virol. 69: 1907-1912, 1995.  
• Fong, S. E., Pallansch, L. A., Mikovits, J. A., Lackman-Smith, C. S., Ruscetti, F. W. and Gonda, M. A.: cis-Acting regulatory elements in the bovine immunodeficiency virus long terminal repeat. Virol. 209: 604-614, 1995.
• Rothblum, C. J., Jackman, J., Mikovits, J., Shukla, R. R., and Kumar, A.: Interaction of nuclear protein p140 with human immunodeficiency virus type I TAR RNA in mitogen-activated primary human T lymphocytes. J. Virol. 69: 5156-5163, 1995.  
• Ouaaz, F., Ruscetti, F., Dugas, B., Mikovits, J., Agut, H., Debré, P., and Mossalayi M. D.: Effects of IgE immune complexes in the regulation of HIV-1 replication and increased cell death of infected U1 monocytes: Involvement of CD23/FceRII mediated nitric oxide and cyclic AMP pathways. Mol. Medicine. 2:1076-1551, 1996.  
• Mikovits, J. A., Hoffman, P. M., Rethwilm, A., and Ruscetti, F. W.: In vitro infection of primary and retrovirus-infected human leukocytes by human foamy virus. J. Virol. 70: 2774-2780, 1996.  
• Hoover, T., Mikovits, J., Court, D., Liu, Y. L., Kung, H. F., and Raziuddin: A Nuclear Matrix-specific Factor that Binds a Specific Segment of the Negative Regulatory Element (NRE) of HIV-1 LTR and Inhibits NF-kappaB Activity. Nucleic Acids Res. 24: 1895-1900, 1996.  
• [Dr. David Daniel Derse (born 1949)], Mikovits, J., Waters, D., Brining, S., and Ruscetti, F.: Examining the molecular genetics of HTLV-1 with an infectious molecular clone and permissive cell culture systems. J. AIDS and Human Retrovirology. 12: 1-5, 1996.  
• Turley, J. M., Fu, T., Ruscetti, F. W., Mikovits, J. A., Bertolette III, D. C., and Birchenall-Roberts, M. C.: Vitamin E succinate induces Fas-mediated apoptosis in estrogen receptor-negative human breast cancer cells. Cancer Res. 57: 881-890, 1997.  
• [Dr. David Daniel Derse (born 1949)], Mikovits, J.A., and Ruscetti, F.W. X-I and X-II open reading frames of HTLV-I are not required for virus replication or for immortalization of primart T cells in vitro. Virology 237:2675-2688, 1997  
• Mikovits, J. A., Taub, D. D., Turcovski-Corrales, S. M., and Ruscetti, F. W.: Similar levels of HIV replication in Th1 and Th2 clones. J. Virol. 72:5231-5238 1998.  
• Mikovits, J. A., Young, H. A., Vertino, P., Issa, J. P. J., Pitha, P. M., Turcoski-Corrales, S., Taub, D. D., Petrow, C. L., Baylin, S. B., and Ruscetti, F. W.: HIV-1 infection upregulates DNA methyltransferase resulting in de novo methylation of the IFN-gamma promoter and subsequent downregulation of IFN-gamma production. Mol. Cell. Biol.,18:5166-5177, 1998.  
• Nilsson, G., J. A. Mikovits, D. D. Metcalf and D.D.Taub.: Mast cell Migratory Response To interleukin-8 Is Mediated Through Interaction With Chemokine Receptor CXCR2/Interleukin-8RB. Blood, 93:2791-2797, 1999.  
• Li, B-Q, Fu T., Dongyan Y., Mikovits JA. Ruscetti FW., Wang JM. Flavonoid Baicalin inhibits HIV-1 infection at the level of viral entry. Biochem Biophys Res Commun. 24: 276(2): 534-8, 2000  
• B.-Q Li, M.A. Wetzel, J.A. Mikovits, E.E. Henderson, T.J. Rogers, W. Gong, N.M. Dunlop, Y. Le, F.W. Ruscetti and J.M. Wang. The synthetic peptide WKYMVM attenuates the function of the chemokine receptors CCR5 and CXCR4 through activation of the formyl peptide receptor-like 1. Blood 2001 May 15; 97(10): 2941-2947.
• M.R. Ruff, L.M. Melendez-Guerrero, Q-E Yang, W-Z Ho, J.A. Mikovits, C.B. Pert and F.W. Ruscetti. Peptide T Inhibits Chemokine Receptor-5 (CCR5) Mediated HIV-1 Infection. Antiviral Research 2001 52: 63-75.  
• J. Mikovits , F. Ruscetti, W. Zhu, R. Bagni, D. Dorjsuren and R. Shoemaker. Potential Cellular Signatures of Viral Infections in Human Hematopoietic Cells. Disease Markers. 2001;17(3):173-8.  
• J-Y Fang, JA Mikovits, R Bagni, CL Petrow-Sadowski and FW Ruscetti. Infection of Lymphoid Cells by Integration-Defective HIV-1 increases De Novo Methylation. J Virol. 2001. 75: 9753-9761.  
• Song Y, Goel A, Basrur V, Roberts PE, Mikovits JA, Inman JK, Turpin JA, Rice WG, Appella E. Synthesis and biological properties of amino acid amide ligand-based pyridinioalkanoyl thioesters as anti-HIV agents. Bioorg Med Chem. 2002 May;10(5):1263-73 .
• Yang QE, Li KG, Mikovits JA. Eradication of human immunodeficiency virus type 1-infected cells by a combination of antimetabolic cytotoxic chemotherapy and antiviral chemotherapy in vitro: a pilot study. J Infect Dis. 2002 Sep 1; 186(5):706-9.  
• Stephen AG, Worthy KM, Towler E, Mikovits JA, Sei S, Roberts P, Yang QE, Akee RK, Klausmeyer P, McCloud TG, Henderson L, Rein A, Covell DG, Currens M, Shoemaker RH, Fisher RJ. Identification of HIV-1 nucleocapsid protein: nucleic acid antagonists with cellular anti-HIV activity. Biochem Biophys Res Commun. 2002 Sep 6; 296(5):1228-37.  
• Flynn J, Fang JY, Mikovits JA, Reich NO. A potent cell-active allosteric inhibitor of murine DNA cytosine C5 methyltransferase. J Biol Chem. 2003 Mar 7;278(10):8238-43. Epub 2002 Dec 10.  
• Muegge K, Young H, Ruscetti F, Mikovits J Epigenetic control during lymphoid development and immune responses: aberrant regulation, viruses, and cancer. Ann N Y Acad Sci. 2003 Mar; 983:55-70. Review  
• Puri A, Rawat SS, Lin HM, Finnegan CM, Mikovits J, Ruscetti FW, Blumenthal R. An inhibitor of glycosphingolipid metabolism blocks HIV-1 infection of primary T-cells.AIDS. 2004 Apr 9;18(6):849-58.  
• Taub DD, Mikovits JA, Nilsson G, Schaffer EM, Key ML, Petrow-Sadowski C, Ruscetti FW. Alterations in mast cell function and survival following in vitro infection with human immunodeficiency viruses-1 through CXCR4. Cell Immunol. 2004 Aug;230(2):65-80.  
• Fang JY, Lu R, Mikovits JA, Cheng ZH, Zhu HY, Chen YX. Regulation of hMSH2 and hMLH1 expression in the human colon cancer cell line SW1116 by DNA methyltransferase 1. Cancer Lett. 2006 Feb 20; 233(1):124-30. 
• Whitby D, Marshall VA, Bagni RK, Miley WJ, McCloud TG, Hines-Boykin R, Goedert JJ, Conde BA, Nagashima K, Mikovits J, Dittmer DP, Newman DJ. Reactivation of Kaposi's sarcoma-associated herpesvirus by natural products from Kaposi's sarcoma endemic regions. Int. J Cancer. 2007 Jan 15;120(2):321-8.  
• Lombardi, V.C., Ruscetti, F. W., Das Gupta, J., Pfost, M.A., Hagen, K.S., Peterson, D.L., Ruscetti, S.K., Bagni, R.K., Petrow-Sadowski, C.L., Gold, B., Dean, M., Silverman, R.H. and J. A Mikovits. Detection of an Infectious Retrovirus, XMRV, in Blood Cells of Patients with Chronic Fatigue Syndrome. Science. 326:585-589, 2009 *retracted by editorial decision 11/2011  
• Mikovits, JA and FW Ruscetti 2010. Response to Comments on “Detection of an Infectious Retrovirus, XMRV in Blood cells of Patients with Chronic Fatigue Syndrome. Science 328, 825-d (2010)  
• Mikovits, JA, Lombardi, VC and FW Ruscetti. Xenotropic Murine Leukemia Virus Related Virus (XMRV): Current Research, Disease Associations, Therapeutic Opportunities (Future Medicine, Therapy, Sept 2010)  
• Mikovits JA, VC Lombardi, MA Pfost, KS Hagen and FW Ruscetti. Addenda to: Detection of an infectious retrovirus, XMRV, in blood cells of patients with chronic fatigue syndrome. Virulence. 2010 Sep-Oct;1(5):386-90. 
• Mikovits JA Huang Y, Pfost MA, Lombardi VC, Bertolette DC, Hagen KS, and Ruscetti FW. Complementary Methods are needed to reveal the extent of distribution of XMRV Infection in Chronic Fatigue Syndrome and Prostate Cancer. AIDS Rev. 2010 Jul-Sep;12(3):149-52. Review.  
• Carruthers BM, van de Sande MI, De Meirleir KL, Klimas NG, Broderick G, Mitchell T, Staines D, Powles AC, Speight N, Vallings R, Bateman L, Baumgarten-Austrheim B, Bell DS, Carlo-Stella N, Chia J, Darragh A, Jo D, Lewis D, Light AR, Marshall-Gradisbik S, Mena I, Mikovits JA, Miwa K, Murovska M, Pall ML, Stevens S. Myalgic encephalomyelitis: International Consensus Criteria. J Intern Med. 2011 Oct;270(4):327-38.  
• Lombardi VC, Hagen KS, Hunter KW, Diamond JW, Smith-Gagen J, Yang W, Mikovits JA. Xenotropic murine leukemia virus-related virus-associated chronic fatigue syndrome reveals a distinct inflammatory signature. In Vivo. 2011 May-Jun;25(3):307-14.  
• Simmons G, Glynn SA, Komaroff AL, Mikovits JA, Tobler LH, Hackett J Jr, Tang N, Switzer WM, Heneine W, Hewlett IK, Zhao J, Lo SC, Alter HJ, Linnen JM, Gao K, Coffin JM, Kearney MF, Ruscetti FW, Pfost MA, Bethel J, Kleinman S, Holmberg JA, Busch MP; Blood XMRV Scientific Research Working Group (SRWG). Failure to confirm XMRV/MLVs in the blood of patients with chronic fatigue syndrome: a multi-laboratory study. Science. 2011 Nov 11;334(6057):814-7. 
• Silverman RH, Das Gupta J, Lombardi VC, Ruscetti FW, Pfost MA, Hagen KS, Peterson DL, Ruscetti SK, Bagni RK, Petrow-Sadowski C, Gold B, Dean M, Mikovits JA. Partial retraction. Detection of an infectious retrovirus, XMRV, in blood cells of patients with chronic fatigue syndrome. Science. 2011 Oct 14;334(6053):176  
• Alter HJ, Mikovits JA, Switzer WM, Ruscetti FW, Lo SC, Klimas N, Komaroff AL, Montoya JG, Bateman L, Levine S, Peterson D, Levin B, Hanson MR, Genfi A, Bhat M, Zheng H, Wang R, Li B, Hung GC, Lee LL, Sameroff S, Heneine W, Coffin J, Hornig M, Lipkin WI. A multicenter blinded analysis indicates no association between chronic fatigue syndrome/myalgic encephalomyelitis and either xenotropic murine leukemia virus-related virus or polytropic murine leukemia virus. MBio. 2012 Sep 18;3(5).

On the uncharacteristically cool morning of July 22, 2009, 24 scientists gathered at the National Institutes of Health in Bethesda, Maryland. The topic: a troubling new retrovirus called XMRV. A paper in the journal Science was about to implicate the virus as the cause of a devastating disease with a dismissive name: chronic fatigue syndrome, or CFS. Doctors have compared the worst cases of CFS to end-stage AIDS. CFS patients have cancer rates that are significantly elevated and immune systems that are seriously impaired. The disease can confer a kind of early dementia and may shorten its victims’ lives by 20 years.

Scientific explanations for the rise of CFS cases, a phenomenon dating to the mid-1980s, have mostly focused on viruses, but psychiatric theories have abounded, too, driven primarily by the Centers for Disease Control and Prevention, which promoted the idea that CFS was “hysteria” or hypochondria. Many scientists and doctors familiar with the disease had long suspected a retrovirus, an organism rife in nature that invades the immune and central nervous systems, as seen in AIDS. Once a retrovirus has infected an organism, it commandeers that organism’s genetic machinery, turning a once-healthy cell into a retroviral powerhouse that spreads the infection to more cells in an irreversible cascade. 

By 2009, scientists had identified a mere handful of specifically human retroviruses: HIV 1 and 2 (human immunodeficiency virus), the cause of AIDS; the HTLV group (human T-cell lymphotrophic virus), at least one of which — HTLV-1 — was a cause of leukemia, lymphoma and an MS-like neurological disease; and HBRV (human beta retrovirus), tentatively considered the cause of a severe liver disease called primary biliary cirrhosis.

At the center of speculation about the new retrovirus that day in July was an immunologist and AIDS researcher named Judy Ann Mikovits (pronounced My-ko-vitz), a diminutive 51-year-old in a sleek black suit and a crisp white shirt. Mikovits was a 20-year veteran of the National Cancer Institute (NCI) who had coauthored more than forty scientific papers. During her final two years at the agency, she had directed the Lab of Antiviral Drug Mechanisms, where she studied therapies for AIDS as well as one of its associated cancers, Kaposi’s sarcoma.

Mikovits had flown to the NIH meeting from Reno where she had been working in a small lab on the campus of the University of Nevada medical school for the previous three years. She listened as other scientists presented what they knew about XMRV, which was largely that it might be linked with prostate cancer. 

Three years earlier, investigators had found genetic sequences of the novel retrovirus in prostate tumors from men with that disease. Whether XMRV caused any human diseases — or was even a transmissible, specifically human infection — was uncertain at best. 

Last to speak, Mikovits led her audience through a terse, rapid-fire slide presentation. She offered persuasive data suggesting that XMRV was infecting a large majority of those suffering from the myriad neurologic and immunologic abnormalities of CFS, thought to afflict between 1 million and 4 million Americans and perhaps 17 million worldwide. Mikovits had found evidence for retroviral infection in blood and plasma (and would later find evidence in saliva). She also offered proof that patients had mounted an immune response to the virus.

Frank Ruscetti, who had discovered HTLV-1 while working in Robert Gallo’s Laboratory of Tumor Cell Biology at the NCI in 1980, was Mikovits’s primary collaborator. It was Ruscetti who had found the new virus in both T-cells and B-cells of CFS patients. Using a labor-intensive cell culturing technique for hunting retroviruses that he had pioneered in the 1970s, Ruscetti had transmitted the pathogen from patients’ T-cells to uninfected T-cells in the laboratory. 

The implications were hardly lost on the Bethesda crowd: If the virus was transmitted in cell cultures in Ruscetti’s lab, it could also be contaminating the nation’s blood supply as a result of blood donations from unknowingly infected donors. As if to confirm those fears, Mikovits reported evidence for XMRV in 3.75 percent of a group of 218 people serving as healthy controls, selected randomly by zip code. Extrapolated to the general population, that meant the recently discovered retrovirus might already have infected at least 10 million still-healthy Americans, each of them innocent of their acquisition of the pathogen. 

Probably the most surprising aspect of what Mikovits said that day, however, had to do with the fact that the new virus appeared to be in the murine leukemia family, which belonged to the larger retrovirus genus known as gammaretroviruses. The latter infect vertebrates ranging from gibbon apes to koala bears to reptiles to domestic cats and are known to cause leukemia, as well as neurological disease. 

Murine leukemia viruses, or MLVs, are carried in the genomes of mice (murine comes from the Latin for “mouse”). MLVs so dependably cause cancer in lab-bred mice — especially leukemia and lymphoma — that a small fraternity of scientists at the NCI and elsewhere has fruitfully studied these viruses since the 1960s in an effort to understand how human cancer begins.

Given such far-reaching implications, it was not surprising that when Mikovits stopped talking, nearly a minute passed before someone spoke, and then it was to say, “Oh my God.”

That simple expression of dread was the preliminary gasp in what would become, in the three years that followed, the most clamorous scramble in recent medical history to prove or disprove what seemed to be a viable hypothesis, one so dire it was facetiously dubbed the Doomsday Scenario by one skeptic. 

Indeed, the controversy over Mikovits’s findings didn’t even have a chance to smolder; it erupted like a wildfire on several continents. The contentious debate reached an apex in the spring of 2010, when one group of government scientists at the FDA found evidence to support Mikovits’s CFS discovery, and another group at the CDC seemed to discredit her findings by failing to reproduce them. 

At stake was the resolution of a potentially infectious disease afflicting millions worldwide — or, in the naysayers’ view, the timely dismissal of a wrongheaded avenue of investigation.

In the end, the NIH’s AIDS czar, Anthony Fauci, asked his friend Ian Lipkin, a neurologist and virus hunter at the Center for Infection and Immunity at Columbia University’s Mailman School of Public Health, to settle the impasse. Last September Lipkin announced the results of his analysis at a tightly controlled press conference: XMRV was not actually a human pathogen, he said, confirming an earlier report, but a man-made contaminant unwittingly manufactured in a lab in the 1990s. 

After novelistic plot twists that included a police raid, an arrest, and a public tarring and feathering of the outspoken Mikovits, the scientific community is now convinced that XMRV was a false lead. Yet the expenditure of time and money to reach that conclusion was hardly an exercise in futility. 

The 2009 meeting in Bethesda marked the beginning of a change in the CFS cosmos, with a slew of prestigious scientists entering the field and government officials taking a more serious view of this formerly neglected disease. Moreover, the XMRV saga opened a rare window into the way scientists and government officials respond to findings that not only challenge the status quo but have profound implications for public health. 

JUDY MIKOVITS HAD BEEN SO COCOONED IN the world of AIDS that she had never heard of chronic fatigue syndrome until 2006, when she was hired to consult for a Santa Barbara–based foundation supporting investigation into human herpes virus six (HHV6), which had been implicated in the disease. At the time, she was newly married and living in California, working on cancer therapies for a small drug-development firm. Mikovits attended the HHV6 Foundation’s annual spring scientific conference, held that year in Barcelona. 

One of the presenters was Daniel Peterson, an internal medicine specialist and an internationally known clinical expert in CFS from Nevada, where a well-publicized outbreak of the disease had occurred in communities near Lake Tahoe in 1984 and 1985. Mikovits listened, fascinated, as Peterson described a group of 300 CFS patients in his practice who suffered from white matter brain lesions, a host of immune system abnormalities, and myriad chronic viral infections including HHV6.

More interesting to Mikovits, however, was Peterson’s report that 5 percent of this group had developed rare immune system cancers during their illness. In all, 77 of the 300 had lymphoma, leukemia, or cellular changes that were predictive of those diseases. Mikovits believed that the signature of the immune system deficiencies described by Peterson, in combination with his report of lymphomas, was strongly reminiscent of the immune dysfunction, opportunistic infections, and cancers of AIDS.

The hypothesis was so dire that one skeptic facetiously dubbed it the Doomsday Scenario.

Also in the audience in Barcelona that day was Annette Whittemore, a rich, politically well-connected Nevada native. Her adult daughter had suffered from CFS since childhood and had been under Peterson’s care for many years. Whittemore and her husband, Harvey — a Reno attorney, real estate magnate, and former lobbyist whom local reporters called “the most powerful man in Nevada” — had been holding fund-raisers at their summer vacation mansion overlooking Lake Tahoe for some years to support research into their daughter’s illness. 

They also had been lobbying the state legislature and U.S. Senator Harry Reid, a longtime friend, for government money to fund an institution at the University of Nevada in Reno dedicated to the study of CFS. In 2005 the state legislature approved a bill committing $12 million to get the $77 million construction project off the ground. (A variety of other sources, including the Whittemores themselves, would fund the rest.) 

Plans were laid for a steel and glass-clad building on the UNR campus that would house, four years hence, the Whittemore Peterson Institute: a collection of labs, clinics, and the office of the president, Annette Whittemore.

Whittemore was impressed when Mikovits — first to the microphone to comment when Peterson finished — said, “I’m a cancer researcher. Number one, I look for viruses in cancer, and number two, this smells like a virus.” Whittemore invited Mikovits to Reno for a six-week stay that summer, during which Mikovits developed a bank of CFS patient tissues for further study. The many patients too weak to stand or walk, requiring wheelchairs or scooters or — to keep their balance — canes, made an impression. By summer’s end she had accepted an offer to become research director of the institute. 

ONE OF THE FIRST THINGS MIKOVITS DID was to employ a microarray — a small tray seeded with DNA from nearly every known virus — to flag viral DNA in human white blood cells. When Mikovits used the microarray to study CFS patients, she “saw all these infections — lots of chronic, active infections — cytomegalovirus, Epstein-Barr virus, human herpes six,” she says. It looked like an acquired immune deficiency syndrome, “like an AIDS patient,” with a weakened immune system enabling opportunistic infections to take root. The question was, why did they have chronic immune dysfunction?

Judy Mikovits stands outside her former lab at the Whittemore Peterson Institute in Reno, Nevada, where she tried to prove a retroviral cause of CFS. (Credit: David Calvert/AP Images)

Mikovits was keenly aware of two well-documented immune system abnormalities in CFS. One was a stark deficiency in the total number of natural killer cells, which seek out and destroy viruses and cancer cells, and a reduction in the killing power of the natural killer cells that remained. 

A second abnormality had to do with elevated levels of RNase L, an enzyme that provides immunological defense against viruses and cancer. “Given that the job of RNase L was to chew up RNA viruses, we hypothesized the possibility of a novel RNA virus in our patients,” Mikovits says. (RNA viruses use ribonucleic acid as their genetic material; examples include SARS, polio, and retroviruses like HIV.)

Patients with aggressive prostate cancer, the second-leading cause of cancer deaths among men, had RNase L abnormalities too. In 2002, Robert Silverman, an RNase L expert at the Cleveland Clinic, discovered that a mutation in the RNase L gene was frequently present in men prone to early onset of the disease. 

Silverman, too, suspected an RNA virus might be the infectious cause of aggressive cancers in relatively young men. And he had some preliminary evidence: When he added DNA from malignant prostate tissue cells to a microarray, a novel retrovirus emerged. Further analysis showed that the retrovirus was a gammaretrovirus closely linked to MLVs found in mice, though analysis revealed its genome to be slightly — less than 3 percent — different. 

Silverman’s group at the Cleveland Clinic and collaborators Joe DeRisi and Don Ganem in San Francisco reported their discovery of a new, apparently human retrovirus in PloS Pathology in March 2006; the researchers added the word xenotropic to the name to indicate that the virus, although carried from generation to generation in the mouse genome, caused disease only in other animals. At the time, the paper attracted little interest from the scientific community.

In January 2009, Mikovits and Ruscetti — by then also hot on the trail of the enigmatic virus — met with Silverman in San Diego, where he had been invited to speak. Over lunch, Mikovits laid out her preliminary data. Using polymerase chain reaction (PCR) technology to greatly amplify what began as tiny amounts of the virus, she had found evidence for the novel pathogen in CFS patients as well. “We all agreed it was worth following up,” Mikovits recalls.

Silverman and Ruscetti signed a confidentiality agreement with the Whittemore Peterson Institute and became Mikovits’s collaborators. Their paper, “Detection of an Infectious Retrovirus, XMRV, in Blood Cells of Patients with Chronic Fatigue Syndrome,” appeared in the online edition of Science on October 8, 2009. Two MLV experts, one from Britain and another from the United States, extolled the importance of the work in an accompanying editorial, writing, “If these figures are borne out in larger studies, it would mean that perhaps 10 million people in the United States and hundreds of millions worldwide are infected.” 

By November 2009, Stuart Le Grice, head of an influential NCI division called the Center of Excellence in HIV/AIDS and Cancer Virology and the person who had convened the NIH meeting the previous summer, had 10 HIV scientists working full time on XMRV. “It’s very interesting to be in the middle of a situation like this,” Le Grice commented at the time. “I worked with Hoffmann–La Roche for seven years in Switzerland in the first days of HIV, and the action plan that we put together at Roche is the same plan that we put together here.”

Mikovits, for her part, took pains to tell members of the lay press and other scientists that the virus had not been proved to cause CFS, merely that it was “associated” with the disease. But in private, contemplative moments that fall, the researcher indicated she felt confident that the long-enduring CFS puzzle was likely solved; her real interest going forward was in developing treatment for those who were suffering.

WHILE SCIENTISTS AT THE NCI WERE repurposing laboratory space to study XMRV, William Reeves, the researcher in charge of CFS for 18 years at the CDC (he died last August) and a longtime proponent of the psychiatric explanation for the disease, sounded a sour note, telling The New York Times, “If we validate it, great. My expectation is that we will not.” 

For many researchers who had followed the CDC’s desultory history in CFS over the years, Reeves’s comments aroused concern. Twenty years earlier, another scientist, immunologist Elaine DeFreitas of the Wistar Institute in Philadelphia, had discovered retroviral gene sequences in CFS sufferers. 

The agency failed to replicate her findings after a series of lab mishaps and a wholesale departure from DeFreitas’s protocol for finding the virus. Afterward, the CDC reverted to its years-long claim that CFS was lacking in biological abnormalities and its assertion to doctors that performing any but the most standard tests was a waste of money. 

Academic research into the cause of the disease went into a 20-year stall, punctuated in 2006 by a press conference in which Reeves and the CDC’s director at the time, Julie Gerberding, announced what they portrayed as a breakthrough discovery: CFS was caused by a genetic predisposition to difficulty responding to stress. 

In late 2009 at CDC headquarters in Atlanta, a virologist in the agency’s retroviral branch, William Switzer, was attempting to replicate the XMRV finding in CFS. In an ironic twist, Switzer was working under the direct supervision of Walid Heneine, the very scientist who had failed to replicate DeFreitas’s work 20 years before. So far, the agency had found XMRV in 2 out of 162 men with prostate cancer but was having zero luck finding it in agency-selected CFS patients.

By the start of 2010, it seemed as if the Science article had launched a cottage industry of researchers on two continents who were unable to find XMRV in CFS patients. The first negative results appeared in January when a group from Imperial College London published an article in PLoS ONE saying they had failed to find the virus in 186 CFS patients using PCR technology. 

In February, a Dutch group at Radboud University followed with another negative PCR study, reporting in the British Medical Journal that it had been unable to find XMRV in 32 patients or 43 healthy controls. Back in the United States the following summer, the CDC reported in the journal Retrovirology its inability to find XMRV in either CFS patients or controls.

Despite the skepticism emerging in Europe and the U.S., events were quietly unfolding in a laboratory at the FDA in Rockville, Maryland, that would put Mikovits’s critics back on the defensive. 

Ian Lipkin of Columbia University orchestrated a grand experiment to learn whether XMRV could be found in patients with CFS. The answer: No. (Credit: Columbia University)

Shyh-Ching Lo, a pathologist and director of the FDA’s Tissue Microbiology Laboratory, discovered that he possessed 37 unopened vials of whole blood and plasma from CFS sufferers, the pristine cryopreserved remains of experiments undertaken in the early 1990s. To his surprise, Lo found four different MLV-related gene sequences in 32 of the 37 patient samples. 

Although the gene sequences were not identical to the Mikovits-Ruscetti XMRV gene sequence reported in Science, they were so close Lo believed he had found genetic variants of a single MLV-like virus species that likely included XMRV. Lo was encouraged by the variants because retroviruses are extremely mutable pathogens that change their gene sequences again and again in response to immune system efforts to kill them. “This is what we expect,” Lo said at the time.

Lo asked a friend and sometime collaborator, Harvey Alter, chief of the infectious disease section in the department of transfusion medicine at the NIH, for some healthy blood samples to use as controls. Alter, who had discovered the hepatitis C virus in the 1970s, provided Lo with blood that had been donated to local blood banks by presumably healthy people. Lo found 6.8 percent of the healthy controls positive for MLV-related infection. The startling figure doubled the Sciencepaper’s estimates of prevalence for Americans who might be infected with a doomsday bug. Lo and Alter’s paper was published in the Proceedings of the National Academy of Sciences in August 2010.

Mikovits and her collaborators were over the moon when they learned of Lo’s success at the FDA. Nevertheless, by the fall of 2010, the majority of published papers were broadly negative. Increasingly, the word contamination was bubbling up in scientific articles and blogs to account for both the XMRV and the MLV-like virus findings. Critics postulated that mouse DNA was floating in the air in the laboratories, or lying in wait in the chemical reagents used to find the virus, or sitting at the bottoms of test tubes used to collect blood specimens.

Le Grice’s influential colleague at the NCI, a government consultant named John Coffin, had his own theory: XMRV had been accidentally created in a lab at Case Western Reserve University in Cleveland sometime between 1993 and 1996. Coffin described how lab workers there had transplanted human prostate tumor cells into an immune-deficient lab mouse, a common procedure for procuring a colony of cells, or a human cell line, for further study. 

When the human cells failed to thrive, lab workers transplanted those same cells into another mouse. Coffin and his collaborator, Vinay Pathak, suggested that with each passage, the human cells acquired genetic portions of a murine leukemia virus, which then merged to form a new virus — a hybrid of the parent sequences. 

The successful prostate cell line that resulted from this event, named “22rv1,” was thus infected with a hybrid, lab-created virus: XMRV. The cell line had been sent to labs around the world — including, as luck would have it, Robert Silverman’s lab at the Cleveland Clinic.

Obviously, if XMRV had been inadvertently manufactured as the result of lab experiments in the mid-1990s, it could not be causing disease in CFS or prostate cancer patients who had fallen ill in the 1980s or early 1990s. XMRV was “dead,” Coffin announced in April 2011. He advised CFS researchers to “move on.”

By early summer 2011, Silverman was ready to accept Coffin’s analysis and had become increasingly uneasy about his contributions to the 2009 data in the Science paper about CFS. Two years earlier, by amplifying the virus in tissue samples, Silverman had sequenced XMRV in seven of 11 CFS patient samples sent to him in March of 2009 by Mikovits. But when he reexamined those original samples again in the summer of 2011, Silverman discovered that all seven were contaminated, not with mouse DNA but with an infectious molecular clone originally made in his own lab in 2006. 

After researchers began using the Silverman clone for their experiments, it replicated “like crazy” and spread beyond the boundaries of those projects throughout the lab, Silverman now found. The discovery was even more troubling since Silverman had sent his synthetic clone to “about a hundred labs” around the world. 

In September, Silverman and the other authors of the Sciencepaper began working with the magazine’s editors on the wording of a partial retraction of the data contributed by Silverman. (Silverman argued for a full retraction.) It was published in the online edition of Science on September 22, 2011, two years after the original study. Bruce Alberts, editor-in-chief of Science, unilaterally retracted the paper in full two months later.

Mikovits nevertheless remained deeply concerned that the prevalence of the Silverman clone in so many laboratories had prevented scientists from looking for greater virus sequence diversity that might have revealed additional MLV-like viruses, as Lo had done at the FDA. No one had yet disproved that a murine leukemia virus was spreading through human population, she insisted—an observation greeted with skepticism and occasional derision by scientists who were following the controversy.

THINGS WERE UNRAVELING BETWEEN ANNETTE Whittemore and Mikovits as well; earlier, on September 29, 2011, Whittemore had fired Mikovits as research director of the Whittemore Peterson Institute. In response to press inquiries, Whittemore offered “insubordination and insolence” as reasons for the dismissal. The dispute had to do with Mikovits’s insistence that another scientist at WPI had improperly ordered a cell line for experiments that were beyond the purview of his authority. Whittemore roundly disagreed with her scientific director.

Three weeks later, Mikovits — who had returned to California to be with her husband — was served with a civil lawsuit by the WPI claiming she had “purloined” lab notebooks and other intellectual property belonging exclusively to the WPI, charges her attorney has vehemently denied. 

Finally, in a development that stunned even her harshest detractors, Mikovits was handcuffed in her home by police on December 4 and booked at the Ventura County jail as a “fugitive from [Nevada] justice.” She remained there for five days, classified as a “flight risk,” until her husband was able to obtain legal representation and have her released on bail.

But L’Affaire Mikovits and the search for retroviral infection in CFS has not come to an end. In January 2012, Ian Lipkin invited Mikovits and the NCI’s Ruscetti to contribute to a rare, NIH-funded study to be orchestrated at Columbia. In fact, Lipkin braved the disapproval of his high-level friends at the NIH to insist that Mikovits be included in the study, which had Ruscetti and Mikovits, the CDC, and the FDA testing blood from nearly 150 CFS patients and an equivalent number of controls. 

Each of the three government teams was given blood samples from the same patients and controls. Lipkin asked each group to employ the lab methods they had used in their original papers. Lipkin and his staff broke the codes and analyzed the results.

None of the participants — including the NCI team — found evidence of XMRV in any samples. Given the definitive results and objective design of the study, the scientific world at last considers the XMRV case closed.

Mikovits is unqualified in her gratitude to Lipkin for insisting that unbiased science be done. XMRV “is one scary virus in the laboratory,” she says today. “It seems to be ultra-stable and can spread through droplets in the air, like a mycoplasma. How could any of us have known?” 

In spite of the disappointing results, Mikovits continues to believe a human retrovirus will be discovered eventually to lie at the heart of CFS. “I still see the footprints of a retrovirus there,” she says. “In the Lipkin study, 3 percent of the controls and 3 percent of the CFS patients had an antibody cross-reactivity to something. It cannot be XMRV, since XMRV doesn’t exist as a human infection. But it’s very close.”

Mikovits was handcuffed in her home by police and booked at the county jail as a ‘fugitive from justice.’ She remained there for five days.

The possibility that Mikovits’s prediction is correct is being explored. A new, $10 million research initiative to investigate CFS was launched in September 2011 at Lipkin’s laboratory after multi-millionaire hedge fund manager Glenn Hutchins, a director of the Federal Reserve Bank of New York and vice chairman of the Brookings Institute, provided the cash and promised more money in the future should the investigation bear fruit. (Lipkin identified the brain infections Borna virus in 1990 and West Nile virus in 1999.) 

“What’s occurred in the last 30 years is criminal,” Mikovits says today. “Mothers and fathers got sick, their children got sick.” But with heightened attention, she adds, patients are likely to get help soon. Even lacking a causal pathogen, biomarkers in this patient population can be studied for clues. “We can find therapies for the CFS patient population even before we determine the exact cause,” Mikovits says. For his part, Lipkin credits Mikovits with “opening Pandora’s box. Right or wrong about this particular virus, she deserves credit for awakening interest in CFS.”

For a disease that has languished in a kind of political never-never land for at least one human generation, leaving millions profoundly disabled, that is significant progress.

The biochemist was fired, then arrested on suspicion of stealing research notebooks — her own research notebooks — from her lab and ultimately shackled to a woman nicknamed Crazy Mary.

She authored a study that turned the science world on its ear, interpreted as a step toward curing a disease, chronic fatigue syndrome, that affects more than 1 million Americans.

In dramas that unfolded as one of the study’s primary conclusions was proved wrong, she lost everything: research opportunities, money, reputation.

Judy Mikovits, sitting in her 38-foot fishing boat in a racket club cap and flip-flops, said through it all she was never given a chance to tell her story.

So she spent weeks and months on this boat, the Pura Vida, in the Channel Islands Harbor, talking, sometimes yelling, into a cellphone. She told of experiences that melded retroviruses named with acronyms with allegations of conspiracy and being blackballed.

The result, “Plague,” written by Mikovits and Kent Heckenlively and released by Skyhorse Publishing, hit bookstores Tuesday.

“I was forced out of science because I found an inconvenient truth,” she said.

GLIMPSE OF HOPE

Before her life slid out of control, Mikovits split time between a laboratory in Reno and the home near Channel Islands Harbor she shared with her husband, David Nolde. She served on a race committee at the Pierpont Bay Yacht Club, volunteering her services as a club bartender.

Known as Judy Nolde, she worked with cancer patients at Community Presbyterian Church in Ventura, where her husband was a church elder.

She was a researcher who said her grandfather’s death to cancer spurred her to a career focused on finding cures. She worked for more than 20 years at the National Cancer Institute, then later at EpiGenX Biosciences in Santa Barbara.

Nearly a decade ago, her studies turned from HIV and cancer to chronic fatigue syndrome, a mysterious, incurable disease marked by exhaustion, pain and memory loss. People with the condition may stay in darkened rooms due to light sensitivity, layering themselves with blankets because their bodies struggle to register warmth.

“I felt the illness had been stigmatized by the scientific community,” said Robin Moulton, a Mikovits supporter from Portland who has struggled with chronic fatigue for 20 years. “ ... There are no tests for diagnosis, treatment or any chance of cure.”

At an institute created by Reno lawyer and former lobbyist Harvey Whittemore and his wife, Anne, Mikovits led a study that offered an explanation for the disease. The work claimed the syndrome was linked to a virus that converts itself into DNA and stays with a person for life. Defined as a retrovirus, it’s called XMRV.

The discovery signaled hope. If the disease was caused or even just linked to a retrovirus, scientists might uncover ways to treat it, maybe cure it.

“All these patients went nuts,” said Vincent Racaniello, a virologist from Columbia University in New York. “They could finally see an end to the suffering.”

The discovery bought acclaim and controversy. Unable to duplicate the studies, some scientists challenged the finding.

Mikovits defended her work. She contended retroviruses could also be linked to autism. Patients from around the world blogged and emailed their support.

“I think they feel she was trying to help this patient population, which has been, like I say, stigmatized,” said Moulton, who dismissed the challenges to Mikovits’ work.

“It just seemed like a lot of negative paper came out quickly,” she said. “It almost seemed like they were trying to take the focus off it.”

GETTING WEIRD

A strange but not unprecedented story of challenged science turned bizarre three years ago on a November Friday.

A woman came to the door of Mikovits’ beachside home outside of Oxnard, claiming to be a patient. Instead, she was a campus police officer from the University of Nevada, Reno. Another Nevada officer came from the bushes. Ventura County Sheriff’s Office deputies emerged from the driveway.

Mikovits was arrested.

She had been fired by the Whittemore Peterson Institute in September over a dispute that involved turning over a cell line to another researcher. After the termination, her laboratory was searched, Mikovits said.

She worried about research notebooks kept in drawers that she was told had been unlocked. They contained her work on XMRV and other studies. They also revealed patient information. She told her assistant to secure the records.

The Whittemores went to the police accusing her of theft. The arrest made national news and led to five days in jail.

“The word I would use is surreal,” Mikovits said. “There’s a 24-hour light in your eye. There’s no pillow. There are no bars. There’s a locked door.”

To this day, she defends her choice to have the notebooks removed from the lab.

“If we had left those notebooks unsecured, patient names would have been exposed,” she said. “It’s like letting your credit card information get out.”

FORCED OUT

She said she lost everything.

The Whittemores sued her. A judge said she was in default because she refused to turn over materials that Mikovits’ lawyer said were protected because of the criminal charges stemming from her arrest.

The criminal case was later dropped. Mikovits filed bankruptcy. Her Oxnard area home was sold. She said the notebooks that triggered her arrest are being held under permanent court injunction, meaning she can’t touch them.

The journal that published her study printed first a partial, then a total retraction. A 2012 study led by [Dr. Walter Ian Lipkin (born 1952)] and also involving Mikovits showed that XMRV was the result of lab contamination.

The study means XMRV isn’t linked to chronic fatigue syndrome, Mikovits said. She also said her research shows a connection to a different, still unidentified retrovirus.

That is the inconvenient truth.

“Retroviruses and environmental toxins led to this explosion of chronic diseases,” she said.

Mikovits wants more research on the retroviral connection not only to chronic fatigue, but cancer, autism and other conditions. She pushes for more studies on the role vaccines and chemicals in the environment play in the diseases.

She doesn’t expect to be involved in the studies or to get a whiff of government research funding.

“She absolutely was blackballed,” said [Dr. Francis William Ruscetti (born 1943)], her research partner in the XMRV study. He said the study and its false conclusion were used to punish Mikovits.

“They did their best to portray it as scientific mischief instead of honest science that didn’t understand the exotic biology of this virus,” he said.

‘PLAGUE OF INTEGRITY’

Mikovits contends leaders of the U.S. Centers for Disease Control and Prevention and the National Institutes of Health saw her work as a threat to their long-held claims the disease was not affected by the environment. She said government officials have long fought efforts to change the way society views chronic fatigue, fearing it would show they covered up what they already knew.

She said her grants from the National Institutes of Health were taken away and given to someone else. She said she was barred from setting foot on the National Cancer Institute, where she once worked.

She said the contaminant that emerged in her study came from a Cleveland lab involved in a study that falsely linked XMRV to prostate cancer. That study wasn’t vilified, Mikovits said, but hers was.

“It’s a plague of integrity,” Mikovits said, explaining her book’s title. “The real plague is the censorship and special interests.”

National Institutes of Health Officials offered no comment on Mikovits’ allegations, saying they support a better understanding of chronic fatigue syndrome. Other observers said they saw no evidence of a conspiracy or government suppression.

“Nobody had anything against Judy,” said Vincent Racaniello, a virologist from Columbia University, citing Mikovits’ efforts to defend her work. “She was simply in the wrong and only at the end did she really admit it.”

Boston researcher John Coffin is described in the book as the grand old man of virology and a supporter of Mikovits’ study who became a foe. He also said he saw no evidence of a conspiracy.

As far as Mikovits’ future, he said he would consider collaborating with her in the right situation.

“I don’t know if she would collaborate with me,” he said.

DESPERATE TO HELP

The story twists more than a tornado. Harvey Whittemore, once Mikovits’ employer, was convicted of funneling $133,000 in illegal campaign contributions to his friend, Sen. Harry Reid, the Senate minority leader. Reid was not accused of wrongdoing.

Whittemore is serving a two-year sentence in federal prison in Lompoc.

Mikovits and her husband live off the pension from his years as personnel director at the Ventura County Health Care Agency. They split time between a small home in Carlsbad and the Channel Islands Harbor. Their boat is up for sale.

She thinks she has been slandered. She probably won’t sue.

“To sue, you have to have a lawyer,” she said. “You have to have money.”

She formed a consulting company with [Dr. Francis William Ruscetti (born 1943)] in Carlsbad. She keeps up to date on research and tries to help the patients who still keep constant contact.

She wants to do more. Moulton, the woman with chronic fatigue, wants the same thing.

“She has so much experience with these things,” Moulton said, referring to cancer, HIV and chronic fatigue. “I think the world would be better with her in a lab.”

This story has been corrected from the original version, which misstated details regarding a search of Mikovits' office. Mikovits said the drawer containing her research notebooks had been unlocked, triggering her request the notebooks be secured.