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D. Kraft, S. C. Joachim, N. Pfeiffer, W. F. Holt, M. B. Wax, F. H. Grus; Antibody Profiles in a Retinal Ganglion Cell Degeneration Model. Invest. Ophthalmol. Vis. Sci. 2007;48(13):2990.
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The aim of this study was to analyze if immunization with HSP27 can cause retinal ganglion cell loss in rats and if changes in serum antibody profiles can be observed.
Lewis rats were immunized with 100 µg HSP27 (plus Freund’s adjuvant and pertussis toxin) and divided into three groups. Rats in group 1 were euthanized after 4 weeks (n=6), in group 2 after 5 weeks (n=6), and in group 3 after 6 weeks (n=9). Group 4: control animals that received no immunization were euthanized after 6 weeks (n=10). Blood was collected from all animals the day they were euthanized. The serum was used to detect IgG antibody patterns against bovine retinal antigens using Western blotting techniques. The antibody patterns were analyzed by multivariate statistical techniques. Eyes were harvested the day the animals were euthanized, all eyes were fixed with 4% paraformaldehyde and the flatmounts were stained with Brn-3a, which detects the nuclei in the retinal ganglion cell layer. A computer assisted quantitation was used to analyze the retinal ganglion cell (RGC) density.
The animals immunized with HSP27 showed a lower number of RGCs in comparison to the control group (P<0.05). All animals, even the control animals, showed complex antibody profiles. Through multivariate statistical methods we could detect a significant difference between group 1 and 4 (P<0.05). Group 2 and 3 showed no significant difference to the control group.
After immunization with HSP27 animals developed complex antibody profiles against retinal antigens. A significant increase in antibody reactivity against HSP27 could be seen in all immunized groups. In previous studies HSP27 was identified as an important antigen in sera of patients with glaucoma. This animal model might be useful to study the development and actions of anti-retinal antibodies and their possible involvement in retinal ganglion cell loss.
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