|Budget Amount *help
¥2,700,000 (Direct Cost : ¥2,700,000)
Fiscal Year 2002 : ¥1,400,000 (Direct Cost : ¥1,400,000)
Fiscal Year 2001 : ¥1,300,000 (Direct Cost : ¥1,300,000)
In this study, to examine abilities of macrophages polarized to oxidative state to suppress experimental autoimmune uveoretinitis(EAU) , we have performed the following experiments.
10 week-old female Lewis rats were immunized with interphotoreceptor retinoid-binding protein emulsified with complete Freund's adjuvant (CFA) to induce EAU, Rats were intraperitoneally injected 20mg/kg of BSO (L-Buthionine-[$, R|-Sulfoximine) to polarize macrophages into oxidative state invivo or PBS asacontrol at 1 2 hrs before and after immunization, atfid at the same time. Besides, BSO was administrated daily from day 0 to day 7 after immunization. Rats were sacrificed day 15 and spleens were collected. Splenic Tcells were cultured in the presence of 1 , 5, 10 ng/ml IRBPfor 4 days. Cultures were pulsed wiith [^3H] thymidine at the last 1 6 hours, followed by cell harvesting and measurement of radioactive. Supernatants were collected from above cultures 48 hours after the initiation, and the concentrations of interferon-y (IFN-y) and interleukin-4 (IL-4) were assessed by enzyme-linked immunosorbent assay. Splenic T cell proliferation responses to IRBPwere consistently lower in BSO-tre&ted rats than that in control rats, whereas their IFN-Y production was observed in BSD-treated rats as well as control rats. IL-4 was not detected in rats treatedwith or without BSO.
In BSD-treated rats, splenic T cell proliferation was significantly decreased, but Th2-type response was not detected. Although we ,changed the doses and administration periods of BSO, the suppressive effects on development of EA!U was not statistically clear.CFA that is used to induce EAU isa strong mediator to polarize macrophages to reductive state. It is unlikely that BSOwould be able to overcome CFA to polarize redox state of macrophages. In future study, spontaneous model thォjtt occur EAU without CFA is required for investigating whether the redox state of macrophage influence EAU development.