Dr. Kathelyn Sue Steimer (born 1948)

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11/10/96 14:10:15 Via Fax

CHIRON

BW)(OBIT/KATHELYN-STEIMER)(CHIR) Kathelyn Sue Steimer of Chiron dies

Business Editors and City Desks

OBIT...

EMERYVILLE, Calif.--(BUSINESS WIRE)--Nov. 18, 1996--Chiron Corp. announced today that Kathelyn Sue Steimer, Ph.D., vice president of research for the company's Chiron Vaccines business unit, died of cancer on Nov. 16 at the age of 48 at her home in Benicia, Calif. Dr. Steimer joined Chiron in 1983 to organize its bioassay group and held a number of research positions, including project leader for Chiron's candidate HIV vaccine, which is in human clinical trials in both the United States and Thailand.

In 1984, together with her colleagues Dr. Dino Dina and Dr. Paul Luciw, Dr. Steimer played a major role in the first sequencing and cloning of the HIV virus, using the SF2 isolate provided by Dr. Jay Levy of the University of California, San Francisco. She then led Chiron's program to research, evaluate and develop genetically engineered proteins of the virus for use in diagnostic assays to detect HIV-1 infection and as candidate HIV vaccines. She also managed the group responsible for evaluating immune responses of humans in clinical trials for Chiron's genetically engineered vaccines for HIV-1, genital herpes, cytomegalovirus and acellular pertussis. As vice president of research, she supervised the vaccines research staff in Emeryville and coordinated their efforts with those of researchers in other vaccine locations throughout the world.

Dr. Steimer served on several National Institutes of Health (NIH) review panels either as an ad hoc or chartered member, and was frequently an invited speaker at national and international scientific meetings. She was an active participant at a number of workshops and special conferences assembled by the National Institutes of Allergies and Infectious Disease, the World Health - Organization and other prominent scientific bodies. -

"At every level of her career, Kathy distinguished herself with - scientific insight, careful analysis and a collaborative approach o that led to productive solutions to important problems," said __ William J. Rutter, Ph.D., chairman and a co-founder of Chiron. "She ___ joined Chiron when we were focused solely on research, helped wit key contributions that created substantial value and opened up avenues of commercial development, then made the transition to managing the efforts and contributions of other scientists and technical staff.

"She was widely appreciated for her contributions advancing understanding of the immune system's response to HIV infection, and in developing the first strong candidate HIV vaccines, the effectiveness of which may yet be proven.

On behalf of Kathy's many colleagues at Chiron but in particular together with its other co-founders Ed Penhoet and Pablo Valenzuela, we thank her for her role in Chiron's growth. All of us will miss her, and will not forget her." Rutter and Penhoet announced the creation of The Kathelyn S. Steimer Scholarship Fund for Graduate Research, which will be used to support academic work leading to a doctoral degree by persons who have worked at Chiron. Persons wishing to add to the fund in Kathy Steimer's memory may send contributions to: The Kathelyn S. Steimer Scholarship Fund for Graduate Research, Chiron Corp., 4560 Horton St., Emeryville, Calif. 94608.

Dr. Steimer earned a B.A. degree in biology from the University of California, Riverside, and a M.S. degree in microbiology from Pennsylvania State University. She then received her Ph.D. in microbiology in 1979 from the University of Pennsylvania. Her graduate research involved studies of the replication and genetics of avian retroviruses in the laboratory of Dr. David Boettiger. Subsequently, she was a postdoctoral research associate at Harvard Medical School in the laboratory of Dr. Judah Folkman, where she was involved in growth factor research, and then was a postdoctoral fellow at Stanford University Medical Center in immunology with Dr. Sam Strober and Dr. Ron Levy, from which she joined Chiron. Dr. Steimer was an author on more than 50 journal articles and contributed to 25 book chapters, many of which were about immunology, viral vaccines and specifically HIV vaccines. She was a principal investigator or co-investigator on six federal or state-funded research grants, all of which studied aspects of HIV infection and vaccines. She was an editorial board member for Journal of Immunotechnology from 1994 on, and for AIDS Research and Human Retroviruses from 1995 to 1996. She was an ad hoc reviewer for Journal of Immunology, AIDS and AIDS Research and Human Retroviruses, and was a member of the American Society for Microbiology.

Dr. Steimer is survived by her husband, Martin Wilson, Professor of Neurobiology, Physiology and Behavior at the University of California, Davis, and their daughter Acacia, both of Benicia, and her father, Harry Steimer, of Sacramento, Calif. The family plans a private ceremony. Chiron will hold a memorial service for colleagues and friends at a time, date and location yet to be determined. Those interested in attending can call Chiron Corporate Communications, 510/923-3309, for more information.

CHIRON CORPORATION * 4560 Horton Street * Emeryville, CA * 94606-2916 * 510-655-8730 * Fax: 510-655-9910

1996 (Nov 24) - NYTimes : "Kathelyn Sue Steimer, 48, AIDS Researcher "

https://www.nytimes.com/1996/11/24/us/kathelyn-sue-steimer-48-aids-researcher.html

1996-11-24-nytimes-kathelyn-sue-steimer-48-aids-researcher.pdf

Dr. Kathelyn Sue Steimer, a leading researcher in the quest for an AIDS vaccine, died on Nov. 16 at her home in Benicia, Calif. She was 48.

The cause was cancer, according to the Chiron Corporation of Emeryville, Calif., where she was vice president for research in the vaccines division.

Dr. Steimer joined the company in 1983 and led a project that developed Chiron's candidate for a vaccine against H.I.V., the virus that causes AIDS. The vaccine is undergoing clinical trials on humans in the United States and Thailand. With Dr. Dino Dina and Dr. Paul Luciw, Dr. Steimer helped bring about the first sequencing and cloning of the virus, in 1985, the company said.

She led a program to evaluate and develop genetically engineered proteins of the virus for use in diagnostic assays. As vice president, she oversaw the vaccine research staff at Emeryville and coordinated efforts with vaccine research centers around the world.

She was a biology graduate of the University of California at Riverside, with a master's degree in microbiology from Pennsylvania State University and a doctorate in that specialty from the University of Pennsylvania.

Dr. Steimer is survived by her husband, Prof. Martin Wilson; a daughter, Acacia, also of Benicia; and her father, Harry Steimer of Sacramento, Calif.

EVIDENCE TIMELINE

1972 (Aug 03) - First wedding

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1987 (April 26) - 

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also - https://www.newspapers.com/image/430760067/?terms=steimer%20chiron&match=1 

1989 book - 

Review : https://core.ac.uk/download/pdf/8219329.pdf

1989-book-review-of-hiv-detection-by-genetic-engineering-methods-luciw-steimer-by-james-chang.pdf

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BOOK REVIEWS

HIV DETECTION BY GENETIC ENGINEERING METHODS. Edited by Paul A. Luciw and

Kathelyn Sue Steimer. New York, Marcel Dekker, Inc., 1989. 301 pp. $99.75.

JAMES CHANG

Medical Student

Yale University School ofMedicine

Technological limitations on human immunodeficiency virus (HIV) detection have left several important questions unanswered. What percentage of infected people actually develop AIDS? Does the patient's antibody response correlate with prognosis? What body tissues does HIV infect? And how is perinatal AIDS transmitted from mother to child? While enzyme-linked immunosorbent assay (ELISA) and Western blot analysis have become established as routine procedures for diagnosis, more sensitive methods crucial for the molecular and epidemiologic dissection of HIV pathogenicity have, until recently, remained primarily research tools. Only now are we able to detect HIV infection by assaying for the virus itself. New methods, notably the polymerase chain reaction, are incredibly sensitive and will allow us to answer those questions concerning AIDS that have so far eluded us.

HIV Detection by Genetic Engineering Methods attempts to bring the reader up to date on these recent breakthroughs. It is a compilation of reviews written by biotechnologists and clinicians, edited by Paul A. Luciw, of the University of California-Davis School of Medicine, and Kathelyn Sue Steimer, of Chiron Corporation. The book's fifteen chapters include all currently used methods for detection of HIV infection, both clinical and experimental. This volume also presents background information on the molecular features of HIV, the epidemiology of AIDS, the genetic variation between HIV- 1 and HIV-2, and the diagnosis of human T-cell leukemia (HTLV) infection.

The middle eleven chapters constitute the heart of this edition. The major development in HIV antibody detection is the use of recombinant HIV antigens and synthetic peptides for serological testing. In the past, ELISA and Western blot tests have relied on purified disrupted virions as the source of antigen. Recombinant HIV antigens and synthetic peptides are preferable for both practical (cost, safety) and scientific (sensitivity, antigen variability) advantages. While these advances in antibody testing are thoroughly discussed, antigen detection methods are relatively ignored. Though there is great excitement over direct detection of the human immunodeficiency virus, the editors devote only four of the fifteen chapters to these techniques. Two chapters review the detection of HIV antigens, and one chapter each is reserved for in situ hybridization and polymerase chain reaction.

HIVDetection by Genetic Engineering Methods lacks emphasis and detail on direct antigen and nucleic acid detection. Each chapter presents only a sketchy outline of the methods and a small sample of results. Precise protocols for these techniques are not included. The references, though extensive, may be found elsewhere quite easily. This book would only be useful to someone who seeks a simple overview of the current state of research in HIV detection. Because this edition is so limited, and because research in this field is advancing at such a dizzying pace, I cannot recommend purchase of this book. 

https://www.nytimes.com/1988/05/15/business/what-s-new-in-vaccines-custom-tailored-for-immunity-without-side-effects.html?searchResultPosition=4

1988-05-15-nytimes-what-s-new-in-vaccines-custom-tailored-for-immunity-without-side-effects.pdf

WHAT'S NEW IN VACCINES

WHAT'S NEW IN VACCINES; 

Custom Tailored' for Immunity Without Side Effects

By Gordon Graff

THE new generation of genetically engineered vaccines, unlike their earlier counterparts, does not contain live or killed disease organisms, which have been linked to side effects. Instead, they make use of only portions of infectious organisms, like surface proteins called antigens, which arouse immunity but seldom cause side effects.

But while gene-splicing techniques would seem to open the door to a flood of new tailor-made vaccines, success of these products is not guaranteed. For one thing, lingering fears of side effects, as well as consumer apathy, could keep demand down. ''All the proof in the world that a vaccine is safe and effective may not matter if people are reluctant to line up for their shots,'' said Mr. Rotheim, the market researcher.

A case in point, he noted, is the hepatitis B vaccine, on the market for two years. The biggest group of recipients of the vaccine, Mr. Rotheim said, have been scientists and health care workers exposed to the hepatitis virus. But intravenous drug users and homosexuals, who are also at high risk for the disease, he said, have been comparatively under-represented among recipients.

Then there is the potential for patent squabbles. In the case of the gene-spliced hepatitis B vaccine, for instance, Biogen, a Cambridge, Mass., biotechnology company, believes its recently awarded patent on certain aspects of the vaccine technology is broad enough to cover production processes used by SmithKline Beckman and Merck. Biogen, in fact, reached a licensing agreement last month with SmithKline Beckman entitling it to continue to make and sell the vaccine in return for royalties.

Regarding Merck, David M. Konys, group product manager of Biogen, said, ''It's obvious they have been infringing on our patent.'' Still, he added, ''it's not inevitable that there's going to be a court battle.'' For Merck's part, a company spokesman said, ''We're confident of our patent position and our ability to continue to sell'' the vaccine. Nonetheless, he too hinted that negotiations are possible, citing Merck's long history of paying royalties to outsiders.

Despite the potential for soft demand and patent disputes, promising new vaccines are still stirring on the horizon. For example, the Chiron Corporation, a small concern that helped develop the hepatitis B vaccine technology used in the Merck product, is trying to bioengineer a vaccine for genital herpes.

Animal tests with the antigen are encouraging, reports Dino Dina, director of virology at Chiron. ''We can not only prevent acute infection but the establishment of chronic infection,'' he says. Right now, Chiron is preparing to apply to the Food and Drug Administration for permission to test its herpes vaccine in humans. If the product is successful, Mr. Dina noted, it would be marketed as a joint venture with Ciba-Geigy.

2007

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https://play.google.com/books/reader?id=mGbMbSZrUTcC&pg=GBS.PP1&hl=en 

Helen Epstein worked direclty for Dr. Kathelyn Steimer

Jitters jeopardize AIDS vaccine trials

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Citation metadata

Author: Jon Cohen

Date: Nov. 12, 1993

From: Science(Vol. 262, Issue 5136)

Publisher: American Association for the Advancement of Science

Document Type: Article 

Length: 1,413 words

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Main content

Abstract: 

Findings reported at the National Institute of Allergy and Infectious Diseases' 6th annual AIDS vaccine conference may delay large-scale efficacy trials of AIDS vaccines. The accuracy of the assay used to determine the effectiveness of the vaccines has been called into question.

AIDS vaccine developers, spurred by the urgency of the pandemic, have been pushing forward - gingerly - with plans for large-scale tests of vaccines. Their goal has been to try experimental preparations in people who are at high risk of becoming infected with HIV, to determine if the vaccines actually can prevent infection. Early lab tests, while they haven't provided any strong leads, haven't thrown up any serious obstacles either: These vaccines appear safe and capable of stimulating immune responses. So the developers have been joined in their drive for efficacy trials by the National Institutes of Health (NIH), which last year revealed plans to begin the tests in December 1993.

But discouraging findings reported last week at an annual AIDS vaccine conference have dealt these researchers a serious setback. In the toughest laboratory test that experimental AIDS vaccines have yet faced - which analyzes a preparation's ability to trigger a key immune response against HIV - almost all have failed. Although the results are preliminary, and some researchers raised questions about the accuracy of the assay used to judge the vaccines, the news caused dismay and confusion among the researchers. "It certainly does make me anxious about going forward with large-scale efficacy trials," says Anthony Fauci, head of the National Institute of Allergy and Infectious Disease (NIAID).

Anxiety at NIAID, the world's largest funder of vaccine research, could portend trouble for its partners in the vaccine effort. The World Health Organization (WHO), which is planning with NIAID to stage efficacy trials in four developing countries, could find itself with different criteria than its Bethesda partner for moving forward. Vaccine manufacturers, who have scaled up their operations to begin such trials, fear that NIAID foot-dragging could jeopardize their investment. "There's a lot riding on trying to resolve this issue," says Duke University's Dani Bolognesi, a prominent AIDS vaccine researcher.

The troubling results, reported last week at NIAID's 6th Annual Conference on Advances in AIDS Vaccine Development in Alexandria, Virginia, centered on a battalion of immune warriors known as neutralizing antibodies. While many antibodies can bind to HIV, only neutralizing antibodies can prevent the virus from infecting cells. To assess whether vaccines can raise neutralizing antibodies, researchers run a laboratory test that mixes blood from a vaccinated person with HIV. If neutralizing antibodies are in the blood, HIV will be locked out of healthy cells. One of the reasons that AIDS vaccine efficacy trials have had a green light until now is because just such neutralizing antibodies have been found in the blood samples taken from volunteers participating in several different HIV vaccine trials.

There is, however, a drawback to this sort of assay: It relies on HIV that was grown on continuous laboratory cell lines, and even though antibodies may protect against this lab-adapted virus they may be ineffective against a real-world strain of HIV. A more realistic test is to use HIV that has been freshly harvested from patients, because these "primary field isolates" are believed to be much closer to the type of HIV that would infect a vaccinated person.

This field isolate test was the focus of three new studies presented at the meeting. "We all had anticipated that [neutralizing antibody] titers evident against the lab strains would translate to field isolates," says Duke's Bolognesi. Unfortunately, the scientists were in for unpleasant surprise. First, Bolognesi's collaborator, Thomas Matthews, reported that he and his co-workers analyzed blood from 40 people given six different vaccines. While most of the samples beat back lab-adapted strains of HIV, not one could neutralized a field isolate. "The obvious implication is that the immunogens used to date don't induce the breadth of response needed to neutralize primary isolates," Matthews said, adding with dark humor: "We hope we don't know what this means."

But the next presentations began to make the meaning depressingly clear. John Mascola of the Walter Reed Army Institute of Research offered another sobering description of how this group also failed to find neutralizing activity against primary isolates when they tested 34 of the blood samples the Duke group analyzed. Mascola was followed by Kathelyn Steimer from California's Chiron Corp., who had focused on people who had received a vaccine jointly being developed by Chiron and CIBA-GEIGY. After testing blood samples on lab isolates, Steimer and co-workers selected the five vaccinees that had the strongest neutralizing antibodies. They then tested these blood samples against primary isolates and, once again, could find no activity.

In workshops and hallway talk, researchers tried to digest the bad news. They debated the vagaries of the neutralizing antibody assays and noted that the differences between the lab-adapted the primary isolates may be the result of everything from laboratory artifacts to different conditions used in the different tests. They also noted that one study presented at the meeting contained hints of success: Barbara Potts from New York's United Biomedical Inc. (UBI) showed that blood from one person injected with UBI's vaccine neutralized a field isolate.

Still, few researchers could dismiss the negative data. "If it turns out that the data is real, any of the products that are presently in clinical trials are not going to be effective," predicts UBI's Wayne Koff, who formerly headed NIAD's AIDS vaccine branch and believes that even the vaccine he is working on needs many more ingredients to be effective. Duke's Matthews thinks action must be taken immediately: "Efficacy trials need to be put on hold right now until this primary isolate question is sorted out."

And that is exactly what NIAID is doing. As John Killen, acting director of NIAID's Division of AIDS, announced at the meeting, NIAID will conduct "a formal and thorough review" next spring of the field isolate issue and decide whether to launch efficacy trials in the United States by the end of 1994.

The hesitation may seem like appropriate scientific caution, but it has infuriated at least one major vaccine developer. Jack Obijeski, head of the AIDS vaccine project at South San Francisco's Genentech, says his company now has more than 200,000 doses of HIV vaccine ready to go because it thought NIAID was committed to moving forward. "To leave that vaccine on the shelf, something that might help someone, we think that's ridiculous," says Obijeski, who doesn't believe the primary isolate question should override other positive animal and human data from experiments with Genentech's vaccine. "If that's the case, this is a monumental disincentive for Genentech.... What needs to be forthcoming is for NIH not to dwaddle about, one step forward, one step back. That's what makes CEOs nervous." He cautions there are many other projects competing with the AIDS vaccine work for company resources.

Jose Esparza, head of AIDS vaccine development at WHO, says developing countries, which will have more than 90% of the world's new HIV infections by 2000, cannot wait for a complete answer to the field isolate question. "Vaccine development is very empirical," says Esparza. "For every point you try to prove there's a counterpoint....I think a trial will give you more information than 1000 lab experiments," WHO, in fact, already is helping Brazil, Uganda, Rwanda, and Thailand prepare for efficacy trials and has run into what he sees as a much more formidable obstacle: Vaccine manufacturers have little interest in tailoring vaccines specifically for these countries - which have different strains of HIV - when the market is uncertain. "There's a need here to encourage manufacturers to make strain-specific vaccines," says Esparza.

The head of the United States military's AIDS vaccine program, Donald Burke, concurs with this view. For more than a year, Burke has been trying to find a company willing to make a vaccine for the strain of virus circulating in Northern Thailand, where he is helping to lay the groundwork with Thai officials for efficacy trials. "I don't want to close the door, but none of the manufacturers has made a commitment up to this point," says Burke.

NIAID's Margaret Johnston ended the conference with the reminder that "none of us are willing to wait for the ideal vaccine" before starting efficacy trials. But if the field isolate findings hold up, expect a heated debate about the risk of jumping in versus the risk of standing still.

Copyright: COPYRIGHT 1993 American Association for the Advancement of Science

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Interview with Dr.Paul Luciw, CCM, University of California, Davis

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Dr.Paul Luciw, Professor, UCDavis is a virologist who cloned the genome of HIV in 1982.  For studies on simian immunodeficiency virus (SIV), a major aim of Dr. Luciw’s research is to identify viral determinants of pathogenesis in the non-human primate model (i.e., rhesus monkeys) for AIDS.


Infection of Cats by Injection with DNA of a Feline Immunodeficiency Virus Molecular Clone

https://pdf.sciencedirectassets.com/272412/1-s2.0-S0042682200X01286/1-s2.0-S0042682297987870/main.pdf?X-Amz-Security-Token=IQoJb3JpZ2luX2VjEBMaCXVzLWVhc3QtMSJHMEUCIQCmr4JasCrL6xLkfJh2x80NqoIhIKeoc1VpaP1V1IRT2QIgbnzVyU85bkaVRIqSu36BkXloq9koSou3Yy8iZowrbQ8qgwQIy%2F%2F%2F%2F%2F%2F%2F%2F%2F%2F%2FARAEGgwwNTkwMDM1NDY4NjUiDFNK6lt2KAMWS7aG2yrXA6jUrNJDOEXtJxTmjDklUO2JyK6MYZAZRyJYecKh02xd7fQpKF05IgDCwAS7Ppm05%2FZtnXhlYAnWUUMSXHheatJZznmLt0car0HiMmP7SdDd2GhuGh6y6Qxa0tAKxAcT3pJHee8E05bJHr%2FW6s9Oi61xU37OFwCnynBMG%2B2MWhn716%2FJj%2BFPciFkt9xE2YTfpZrcqEADBvlpgfi0l1khn2nVYzqmfj1oFW1E%2FzQq4eEwtqNB7ztadGY3%2FimgefTrB3S3p3H%2FKkufSLqdfpKYY8AuC%2BPS8Y1h%2FvfJL8dOtjE47lrK1r9scafxBspz0aDpbPxiyMfl6WIL%2B6PtOr8efOvqNzwOz61ZI8L4oxMUmeSBHSbFKREzpFOrdx3WZnR7XfOMT63ss%2FQxs35eeCB3VtDWC8J3YP8fU7vxNW1M2NNi40mQaQpnnZUzWesEIrwvdUAPSPORUnkvK3DXxF6eOV%2FblNQNKVQQqPF%2BPmDCg4kghuRdHlwcnNaA7A6eFTPF8WgZ5bhoLk0MDdKpYWUnbJYhWxcPujLo315eoPpGmakXP%2FewBxmf3yKqFmcUwUEMBLmdwIDL23APg2jYFo5Tg%2FzskFQ4ZxGsLBkZD%2FUcmQo6zW59nrtLfDDokdyMBjqlAYoZrVYX5f%2FDst3z4fbudHwpZ%2F0mHr3KA3NFURzrIV7FZxQBcB1kVzCoJ6c%2FKGBlSmIxNvl3uD6CGfWFMqOQ18fklNF4Hgsy%2Fymd5%2FtncnamR01kCKMGJ3C%2FnwZJ7zls2G%2B0HU7X9xGWhBbsCzcppA5we%2B2bki6UVE6YxDnj9McZDdAbzHQ%2F6sdPq%2BqSnHKHxOZV6%2Fz4WHM7ancdfViK5VPT9gkoJA%3D%3D&X-Amz-Algorithm=AWS4-HMAC-SHA256&X-Amz-Date=20211119T022532Z&X-Amz-SignedHeaders=host&X-Amz-Expires=300&X-Amz-Credential=ASIAQ3PHCVTY345XVDCV%2F20211119%2Fus-east-1%2Fs3%2Faws4_request&X-Amz-Signature=de8f03b05b82d96426c8a80aa07af8ef18b77884b319761b63b06ac0eb10a0a4&hash=1ca45f34fb4d2982d77306312b5c7980207609565ed8b9685860617c6aa5eb60&host=68042c943591013ac2b2430a89b270f6af2c76d8dfd086a07176afe7c76c2c61&pii=S0042682297987870&tid=spdf-fed04d30-0251-4c80-baa3-20c13e7963e0&sid=13b96e689c437743e079f3f2557bbbf92460gxrqa&type=client

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VIROLOGY 238, 157–160 (1997) ARTICLE NO. VY978787

BRIEF REPORT

Infection of Cats by Injection with DNA of a Feline Immunodeficiency Virus Molecular Clone

Ellen E. Sparger,*,1 Harry Louie,* Ann Marie Ziomeck,† and Paul A. Luciw† *Department of Medicine and Epidemiology, School of Veterinary Medicine, and Department of Medical Pathology,

School of Medicine, University of California, Davis, California 95616

Received March 24, 1997; returned to author for revision May 28, 1997; accepted August 13, 1997

Establishment of infection of animals with a viral clone will be important for investigating viral determinants of pathogenesis and monitoring sequence changes in the viral genome in vivo and may find utility as a means of immunization with live- attenuated virus. To test the efficiency of intramuscular (im) injection of cloned proviral plasmid DNA for establishing feline immunodeficiency virus (FIV) infection in specific pathogen-free (SPF) cats, groups of cats were inoculated by the im route with 300, 100, or 30 mg of plasmid DNA containing the infectious molecular clone, FIV-pPPR. A fourth group of cats was inoculated intradermally with 30 mg of FIV-pPPR plasmid DNA. For comparison, a fifth group received 103 TCID50 of a live virus stock of FIV-pPPR by intraperitoneal inoculation. Inoculation by im injection with 100 to 300 mg of infectious FIV-pPPR proviral DNA produced infection detectable by both antiviral antibody and virus isolation from peripheral blood mononuclear cells. Inoculation by im injection with 30 mg of proviral DNA resulted in infection in two of three inoculated cats. Intradermal injection with 30 mg of proviral DNA induced infection in one of three cats. Induction of antiviral antibody and viremia was delayed in cats inoculated with 30 mg compared to cats inoculated with either 100 or 300 mg of proviral DNA. This study indicates that cloned FIV proviral DNA may replace infectious virion preparations as inocula for pathogenesis and immuniza- tion studies. 􏰉 1997 Academic Press

 


ArticlePDF Available

Dexamethasone enhancement of gene expression after direct hepatic DNA injection

DOI:10.1016/S0021-9258(18)43966-X

Authors:

Robert Malone

M A Hickman

Karin Bruinsma

T R Sih

Rosemary L Walzem

D M Carlson

J S Powell

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Abstract

The critical physiological functions of the liver make hepatocytes important targets for therapeutic gene delivery. This study reports significant gene expression following direct injection of plasmid DNA into the livers of rats and cats. Transfection was characterized using luciferase and Lac Z expression from plasmids with the cytomegalovirus immediate early promoter/enhancer (CMV IE) or the Rous sarcoma virus long terminal repeat (RSV LTR). Dexamethasone treatment enhanced and prolonged transfected gene expression, possibly by activating gene expression. Southern analysis of total DNA extracted from liver at various times following injection detected persistent unintegrated plasmid DNA which maintained a prokaryotic methylation pattern. This study demonstrates the feasibility of direct DNA injection in the experimental analysis of hepatic gene expression in vivo.

10.1016/S0021-9258(18)43966-X

malone1994.pdf

"Acknowledgments-We thank Karl Mack for the kind gift of pCMvL,

and Suresh Subramani for pRSVL. We also thank Peter Bany and Paul

Luciw for helpful discussions about CMV and viral latency, and Philip

Schneider for assistance with the initial rat surgery."