Articles

Adibhatla, R. M., & Hatcher, J. F. (2008). Phospholipase A(2), reactive oxygen species, and lipid peroxidation in CNS pathologies. BMB reports, 41(8), 560–567.       https://doi.org/10.5483/bmbrep.2008.41.8.560

Back, S. A., Tuohy, T. M. F., Chen, H., Wallingford, N., Craig, A., Struve, J., . . . Sherman, L. S. (2005). Hyaluronan accumulates in demyelinated lesions and inhibits oligodendrocyte progenitor maturation. Nature Medicine, 11(9), 966-72. doi:https://doi.org/10.1038/nm1279 

The study looked specifically at Hyaluronan and its role in Multiple Sclerosis and other degenerative myelin diseases such as EAE. At the lesion sites of MS patients, promyelinating oligodendrocytes and oligodendrocyte progenitors (OPCs) were found. They had not matured enough to myelinate axons. Along with OPCs, hyaluronan was accumulated. The staining intensity of hyaluronan correlated with the level of demyelination in Cnp1-Cd44 mice. In Cnp1-Cd44+/– mice, a slight increase in hyaluronan staining compared to wild-type mice was accompanied by limited evidence of demyelination. In Cnp1+/+ mice, extensive demyelination was accompanied by widespread, intense hyaluronan staining. It was found that Cnp1-Cd44 mice have increased numbers of immature (e.g., O4+MBP–) OPCs throughout adult white matter compared to wild-type mice35. Chronically elevated CD44 expression, accompanied by hyaluronan accumulation, may therefore influence OPC maturation. In early lesions, this hyaluronan probably originates from activated T cells and microglia, which produce smaller forms of hyaluronan that have been thought to be involved in promoting T-cell activation. In older, chronic lesions, a HMW form of hyaluronan is probably synthesized by astrocytes. 

Baeten, K., Adriaensens, P., Hendriks, J., Theunissen, E., Gelan, J., Hellings, N., & Piet Stinissen. (2010). Tracking of myelin-reactive T cells in experimental autoimmune encephalomyelitis (EAE) animals using small particles of iron oxide and MRI. NMR in Biomedicine, 23(6), 601–609. https://doi.org/10.1002/nbm.1501

This study demonstrates the feasibility of tracking SPIO labeled myelin-reactive T cells in the spinal cord and the brain of EAE rats upon systemic administration. Furthermore, the experiment provided data on the optimal labelling conditions for T cells leading to a high particle uptake and minimal aggregate formation.

Billiau, A., & Matthys, P. (2001). Modes of action of Freund's adjuvants in experimental models of autoimmune diseases. Journal of leukocyte biology, 70(6), 849–860. 

Freund's adjuvants are essential for inducing autoimmune disease in animal models. While their precise mechanisms are not fully explored, it is generally believed that incomplete Freund's adjuvant (IFA) and complete Freund's adjuvant (CFA) extend the lifespan of injected autoantigens, enhance their delivery to the immune system, and stimulate the innate immune system, leading to altered immune responses. Early effects include dendritic cell uptake of adjuvant components, increased phagocytosis, cytokine secretion, and CD4+ lymphocyte activation. CFA's mycobacterial components direct T lymphocytes toward a Th1 profile, promoting strong delayed-type hypersensitivity to autoantigens. The mycobacterial elements also induce changes in the hematopoietic system, which can either exacerbate or mitigate autoimmune diseases. In summary, Freund's adjuvants play a pivotal role in both the early and late phases of autoimmune disease induction through their impact on immune responses and hematopoietic changes.

Castro, H. J., Mendez-Inocencio, J., Omidvar, B., Omidvar, J., Santilli, J., Nielsen, H., J., Pavot, A. P., Richert, J. R., & Bellanti, J. A. (2005). A Phase I Study of the Safety of Honeybee Venom Extract as a Possible Treatment for Patients with Progressive Forms of Multiple Sclerosis. Allergy and Asthma Proceedings, 26(6), 470-6 

Castro and their team performed a small study on the effects of bee venom on nine patients with MS. Patients with primary or secondary progressive MS, who were between 21 and 55 years of age, with Kurtzke Expanded Disability Status Scale scores between 4.5 and 8.0 and who did not have other significant medical illnesses were eligible for the study. Over the course of 17 weeks, the patients were injected with western honey bee venom in varying amounts over varying time periods (two week differences). At the end of the study, no conclusive evidence was found to support the use of bee venom on multiple sclerosis in its raw form. 

Constantinescu, C. S., Farooqi, N., O’Brien, K., & Gran, B. (2011). Experimental autoimmune encephalomyelitis (EAE) as a model for multiple sclerosis (MS). British Journal of Pharmacology, 164(4), 1079–1106. https://doi.org/10.1111/j.1476-5381.2011.01302.x

‌This study aimed to compare EAE with MS and describe why EAE is a reliable model for MS in animals. It also mentioned the current in-circulation drugs for MS patients to help relieve symptoms.

Gerrits, P. O., Brekelmans-Bartels, M., Mast, L., s-Gravenmade, E. J., Horobin, R. W., & Holstege, G. (1992). Staining myelin and myelin-like degradation products in the spinal cords of Chronic Experimental Allergic Encephalomyelitis (Cr-EAE) rats using Sudan Black B staining of glycol methacrylate-embedded material. Journal of Neuroscience Methods, 45(1-2), 99–105. 

A high-resolution light-microscopical technique (HRLM) is described to visualize myelin, and macrophages containing degradation products of myelin, in the spinal cords of chronic relapsing experimental allergic encephalomyelitis rats. This HRLM technique was developed to optimize the correlation between nuclear magnetic resonance characteristics and histopathological images in this well-established animal model for MS. Spinal cords were fixed by perfusion with a combination of cacodylate-buffered glutaraldehyde and formaldehyde, post-fixed in Dalton's fixative (containing osmium tetroxide), rinsed in water, processed in ethanol, acetone, and embedded in glycol methacrylate resin (Technovit 7100/HistoResin). Semi-thin sections were stained with Sudan Black B and counterstained with Cresyl Fast Violet, resulting in black staining of myelin and its degradation products, with blue/violet staining of demyelinated axons and other tissue elements.

Ji, Z., Wu, S., Xu, Y., Qi, J., Su, X., & Shen, L. (2019). Obesity Promotes EAE Through IL-6 and CCL-2-Mediated T Cells Infiltration. Frontiers in Immunology, 10. https://doi.org/10.3389/fimmu.2019.01881

In this study, they showed that high fat diet (HFD)-induced obese mice developed an exacerbated EAE as indicated by higher clinical scores and more severe pathological changes in spinal cord than the control mice fed with normal diet (ND), following immunization with myelin oligodendrocyte glycoprotein (MOG) 35–55 peptide. The exacerbation of EAE in HFD mice was associated with enhanced microglial activation and increased expansion of Th1 and Th17 cells. The HFD mice also showed aggravated disease in an adoptive T cell transfer EAE model.

Karimi, A., Ahmadi, F., Parivar, K., Nabiuni, M., Haghighi, S., Imani, S., & Afrouzi, H. (2012). Effect of honey bee venom on lewis rats with experimental allergic encephalomyelitis, a model for multiple sclerosis. Iranian journal of pharmaceutical research : IJPR, 11(2), 671–678. 

The study looked at bee venom and its effects on a common model for multiple sclerosis: experimental allergic encephalomyelitis. Lewis Rats were given 1 mg/ml EAE through Mycobacterium tuberculosis. Hematoxylin and eosin and luxol fast blue methods were used for the analysis of inflammation and demyelination in CNS. Symptoms were recorded based on clinical scores of three groups: saline, E-BV1 (received 2 mg/Kg honeybee venom every day), and E-BV2 (received 5 mg/Kg honeybee venom every day). Both E-BV groups had statistically significant lower average clinical scores when compared to the saline group, with E-BV2 having lower clinical scores than E-BV1. Demyelination showed a decrease for the treatment groups. Demyelination was not observed in the control group, but this process was considerably increased in the saline group. In previous studies, the immune suppression and anti-inflammatory effects of Bee venom have been reported in MS disease, rheumatoid arthritis and their laboratory models. In the EAE rat model, the serum level of TNF-α is increased in the beginning of disease, which shows that TNF-α has an important role in the distribution of disease. During the present study, it was observed that TNF-α is decreased in a group treated with 5 mg/Kg/day of Bee venom. Bee venom can prevent the production of pro-inflammatory cytokines like TNF-α.

Kemeny, D. M., Dalton, N., Lawrence, A. J., Pearce, F. L., & Vernon, C. A. (1984). The purification and characterisation of hyaluronidase from the venom of the honey bee, Apis mellifera. European Journal of Biochemistry, 139(2), 217–223. https://doi.org/10.1111/j.1432-1033.1984.tb07997.x

Hyaluronidase catalyzes the hydrolysis of the long chain mucopolysaccharide, hyaluronic acid, which is important constituent of animal and plant cell walls found in human skin, synovial fluid, tumors, and in group c streptococci. The purpose of the study was to purify hyaluronidase to a high degree for use in the radioallergosorbent test. Previously, the compound was known as unstable. However, in this article, it was found to be extremely stable at storage 4 Celsius. 30% of crude bee venom was found to be hyaluronidase. Using the presence of IgE antibodies to indicate sensitisation IgEd antibodies to phospholipase A were detected in the majority of bee sting allergic patients (91%). 51% of these patients possessed IgE antibodies to hyaluronidase. However, the highest levels of IgG antibodies were to phospholipase A2 and not hyaluronidase.

Krause, T. L., & Bittner, G. D. (1990). Rapid morphological fusion of severed myelinated axons by polyethylene glycol. Proceedings of the National Academy of Sciences, 87(4), 1471–1475. https://doi.org/10.1073/pnas.87.4.1471

Krause and Bittner were able to discover a technique to morphologically fuse severed myelin sheaths in an invertebrate axon. They used the medial giant axon (MGA) of the earthworm Lumbricus terrestris with polyethylene glycol (PEG) to close the cut ends. The fusion rates found could be as high as 80-100%. However, this is only when combined with hypotonic saline and reduced calcium. It is even proposed that the technique may be able to repair mammalian-myelinated axons. On the down side, it is possible that the trauma of the PEG application could disrupt intramembrane particles, cytoskeleton organization, internal calcium concentrations, ATP levels, and axonal-glial relationships.

Nagy, N., Kuipers, H. F., Marshall, P. L., Wang, E., Kaber, G., & Bollyky, P. L. (2019). Hyaluronan in immune dysregulation and autoimmune diseases. Matrix Biology, 78-79, 292–313. 

The scientists described what is known about the size, amount, and distribution of HA at sites of autoimmunity and in associated lymphoid structures in type 1 diabetes, multiple sclerosis, and rheumatoid arthritis. They also examined the recent literature on HA and its impact on adaptive immunity, particularly in regards to the biology of lymphocytes and Foxp3+ regulatory T-cells (Treg), a T-cell subset that maintains immune tolerance in healthy individuals. 

Nazaninalsadat Seyed Khoei, Atashpaz, S., Kamyar Ghabili, Nazlisadat Seyed Khoei, & Yadollah Omidi. (2009). Melittin and hyaluronidase compound derived from bee venom for the treatment of multiple sclerosis. Iranian Journal of Medical Hypotheses and Ideas, 3(3).

The authors looked at previous studies to compile a possible explorational study on the effects of bee venom on Multiple Sclerosis. They believe that the hidden mechanism illustrating the inefficacy of BV in the treatment of MS is phospholipase A2 (PLA2) found in BV. Blockage of PLA2 may hinder onset and progression of MS. A BV component purified of PLA2 may enhance its efficacy of MS treatment. Hyaluronidase also triggers remyelination by degradation of hyaluronan that accumulates in inflammatory demyelinating legions in the CNS of MS patients.

Papadopoulos, D., Pham-Dinh, D., & Reynolds, R. (2006). Axon loss is responsible for chronic neurological deficit following inflammatory demyelination in the rat. Experimental Neurology, 197(2), 373–385. https://doi.org/10.1016/j.expneurol.2005.10.033

In this study, they investigated the features of axonal loss in myelin autoimmunity and tested the hypothesis that loss of axons determines permanent neurological impairment in a model of inflammatory demyelination that closely mimics the pathology and course of MS. EAE was induced in DA rats by injection of recombinant mouse MOG with IFA. Animals that developed progressive EAE were killed at several time points after disease onset and animals that followed a chronic relapsing–remitting course of EAE were killed at approximately 4 months, exhibiting varying degrees of residual disability. Toluidine blue staining of semithin sections and immunohistochemistry for OX-42 were used to quantify demyelination, remyelination, inflammation and axonal loss in the spinal cord of MOG-EAE rats.

Raghuraman, H., & Chattopadhyay, A. (2007). Melittin: a Membrane-active Peptide with Diverse Functions. Bioscience Reports, 27(4-5), 189–223. https://doi.org/10.1007/s10540-006-9030-z

Melittin is the principal toxic component in the venom of the European honey bee Apis mellifera and is a cationic, hemolytic peptide. It is composed of 26 amino acid residues and is primarily hydrophobic. There is vast research on the component as it has a wide varying degree of reactions with lipid-proteins, specifically in their membranes. The study highlighted Melittin's membrane properties using biophysical approaches. 

Robinson, A. P., Harp, C. T., Noronha, A., & Miller, S. D. (2014). The experimental autoimmune encephalomyelitis (EAE) model of MS. Handbook of Clinical Neurology, 173–189. https://doi.org/10.1016/b978-0-444-52001-2.00008-x

EAE is an animal model of autoimmune-mediated inflammatory demyelination that resembles the pathology and symptoms of MS in many ways, but also differs significantly in many aspects of the disease. Much of our understanding regarding the autoimmune and inflammatory process in general comes directly from this model. The successful development and examination of therapies for MS demonstrate the utility of EAE as a model for MS.

Rodriguez-Acosta, A., Vega, J., Finol, H. J., & Pulido-Mendez, M. (2003). Ultrastructural alterations in cortex of adrenal gland caused by the toxic effect of bee (Apis mellifera) venom. Journal of submicroscopic cytology and pathology, 35(3), 309-314. 

Rodriguez-Acosta and their team studied the toxicity of A. Mellifera venom on the adrenal gland. They injected mice with extracted venom and studied its effects 6, 48, and 144 hours after injection. Multiple effects were found including loss of smooth ER, increased fenestrae areas, capillary basal membrane interruptions, as well as new myelin-like figures and autophagic vacuoles. Specifically, the capillary damage resulted in a direct mortal lesion of the endothelium. It could reasonably be suggested that the intervention of histamine and other amines that are found in bee venom take place in the adrenal gland.

Schneider, C., Schuetz, G., & Zollner, T. M. (2009). Acute neuroinflammation in Lewis rats — A model for acute multiple sclerosis relapses. Journal of Neuroimmunology, 213(1-2), 84–90. https://doi.org/10.1016/j.jneuroim.2009.05.015

STOSIC-GRUJICIC, S., RAMIC, Z., BUMBASIREVIC, V., HARHAJI, L., & MOSTARICA-STOJKOVIC, M. (2004). Induction of experimental autoimmune encephalomyelitis in Dark Agouti rats without adjuvant. Clinical and Experimental Immunology, 136(1), 49–55. https://doi.org/10.1111/j.1365-2249.2004.02418.x

Experimental autoimmune encephalomyelitis (EAE) is a well-recognized model for multiple sclerosis (MS) in humans. However, adjuvants used with encephalitogens to induce EAE produce non-specific effects interfering with the mechanisms involved in the autoimmune response to the central nervous system (CNS) tissue. It is therefore important to establish a more suitable model of EAE for analysis of autoimmune phenomena resembling those operative in MS. Here the scientists report that EAE can be induced regularly in Dark Agouti (DA) strain of rats with spinal cord tissue without any adjuvant, as judged by both clinical and histological parameters.

Thessen Hedreul, M., Gillett, A., Olsson, T., Jagodic, M., & Harris, R. A. (2009). Characterization of Multiple Sclerosis candidate gene expression kinetics in rat experimental autoimmune encephalomyelitis. Journal of Neuroimmunology, 210(1-2), 30–39. https://doi.org/10.1016/j.jneuroim.2009.02.010

This article looked at and characterized a Multiple Sclerosis model in rats called Experimental Autoimmune Encephalomyelitis (EAE). The major genetic risk factor in both EAE and MS is the major histocompatibility complex (MHC). Female rats were anesthetized with isoflurane and injected subcutaneously with a 200 µl inoculum containing 15 µg MOG in phosphate buffered saline (PBS), emulsified 1:1 with incomplete Freund's adjuvant at the tail base. After injection, the rats were observed daily for clinical signs of EAE development. PCR was used with SYBR primers to look at cDNA expression. The expression of proinflammatory molecules subsided in the lymph node by day 12 when spinal cord infiltration occurs, thereby initiating EAE in susceptible rats. Hence a combination of TH1 and TH17 responses and macrophage activation leads to neuroinflammation in susceptible rats.

Wang, S.-Z. ., Dulin, J., Wu, H., Hurlock, E., Lee, S.-E., Jansson, K., & Lu, Q. R. (2006). An oligodendrocyte-specific zinc-finger transcription regulator cooperates with Olig2 to promote oligodendrocyte differentiation. Development, 133(17), 3389–3398. https://doi.org/10.1242/dev.02522 

Wang and his coworkers looked at and categorized Zfp488, an oligodendrocyte-specific zinc-finger transcription regulator, by screening for genes downregulated in the optic nerves of Olig1-null mice. They were able to find through data that Zfp488 affects differentiation in oligodendrocytes and their maturation. The scientists harvested brains, spinal cords and optic nerves from wild type and Olig1mutants at various embryonic and postnatal stages. The oligodendrocytes were transfected externally and studied in chick embryos of varying maturities. Zfp488 is initially expressed as foci in the ventral domain of the spinal cord at E14.5, which coincides with the expression of Mbp and Plp1/DM20, two other important aspects to myelination. Zfp488 expression was not seen in peripheral myelinating Schwann cells in the dorsal root ganglion, as expected. To determine whether Zfp488 has counterparts in other species, a homology search revealed that murine Zfp488 appears to have an ortholog but no obvious paralogs in species, including human, rat, chicken and Drosophila. This suggests a conserved non-redundant function for Zfp488 during evolution. The structural feature with two zinc-finger motifs flanking an NLS is highly conserved across all species.

Williams, R. G. (1955). The effects of continuous local injection of hyaluronidase on skin and subcutaneous tissue in rats. The Anatomical Record, 122(3), 349–361. https://doi.org/10.1002/ar.1091220307

The purpose of these experiments was to determine the extent to which the structure, particularly the vasculature, of skin and subcutaneous tissue, could be altered locally by continuous subdermal injections of hyaluronidase in minute amounts.