Oxidat ive macrophage suppresses experimental autoimmune uveoretinitis

氧化巨噬细胞抑制实验性自身免疫性葡萄膜视网膜炎

• 批准号: 13671852
• 负责人: USUI Masahiko
• 金额: $ 1.73万
• 依托单位: Tokyo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671852/
• 关键词: uveit is; oxidative; reductive; experimental autoimmune uveoretinitis; macrophage; Th1; Th2; 抗原提示細胞; レドックス

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.

本研究为了检测巨噬细胞极化至氧化态抑制实验性自身免疫性葡萄膜视网膜炎(EAU)的能力，我们进行了以下实验。用完全弗氏佐剂(CFA)乳化的光感受器间视黄醇结合蛋白免疫10周龄雌性Lewis大鼠以诱导EAU，腹腔注射 20mg/kg BSO（L-丁硫氨酸-[$，R|-磺肟]）使巨噬细胞极化至体内氧化状态，或在免疫前后1-2小时用PBS作为对照，同时注射。此外，从免疫后第0天到第7天每天施用BSO。第15天处死大鼠并收集脾脏。将脾T细胞在1、5、10ng/ml IRBP存在下培养4天。在最后1-6小时用[^3H]胸苷脉冲培养物，然后收获细胞并测量放射性。起始后48小时从上述培养物中收集上清液，并通过酶联免疫吸附测定法评估干扰素-γ（IFN-γ）和白细胞介素-4（IL-4）的浓度。 BSO 治疗大鼠的脾 T 细胞增殖反应对 IRBP 始终低于对照大鼠，而在 BSD 治疗大鼠和对照大鼠中观察到其 IFN-Y 产生。在用或不用BSO治疗的大鼠中均未检测到IL-4。在BSD治疗的大鼠中，脾T细胞增殖显着降低，但未检测到Th2型反应。尽管我们改变了BSO的剂量和给药周期，但对EA!U发展的抑制作用尚不明确。用于诱导EAU的CFA是使巨噬细胞极化至还原态的强介质。 BSO 不太可能克服 CFA 来极化巨噬细胞的氧化还原状态。在未来的研究中，需要建立没有CFA的自发模型来研究巨噬细胞的氧化还原状态是​​否影响EAU的发展。