抗原特异性Th17细胞分化及调节.doc

抗原特异性Th17细胞分化及调节 　 作者：杨静, 贾磊, 李丽, 吴长有 【摘要】 目的: 探讨影响抗原特异性Th17细胞分化和调节的因素。方法: T细胞受体转基因小鼠(DO11.10)CD4+T细胞, 与同背景正常小鼠的脾细胞混合, 经OVA多肽刺激后, 在不同的Th17极化的培养条件下, 观察Th17细胞及细胞因子的产生。 结果: 单纯OVA抗原刺激主要诱导特异性Th1型反应, 在TGFβ和IL6存在的条件下, 主要诱导Th17反应; 当加入IL23之后, 促进了Th17细胞的分化。阻断了IFNγ和IL4之后, Th17细胞的百分率明显增加。LPS可以促进Th1型细胞因子的产生, 但对抗原特异性Th17细胞的分化没有明显的促进作用。结论: 抗原特异性Th17细胞是与Th1、 Th2相对独立的细胞亚群, TGFβ、 IL6和 IL23可诱导或促进Th17的分化, 而Th1和Th2细胞因子抑制其分化。 【关键词】 Th17 T细胞亚群 流式细胞术 细胞因子 　　[Abstract] AIM: To assess the differentiation and regulation of antigen specific Th17 cells in vitro. METHODS: Nave CD4+ T cells from OVATCRtransgenic mice (OVATCRTg) were isolated and cocultured with splenocytes from normal BALB/c mice under different culture conditions. The expression and production of IL17 and other cytokines were assessed. RESULTS: OVA peptide alone mainly induced Th1 type responses. Addition of TGFβ plus IL6 induced Th17 response. Furthermore, addition of IL23 to cell culture could promote the differentiation of Th17 cells. Blocking of IFNγ and IL4 by neutralizing monoclonal antibodies enhanced the percentage of IL17producing cells. LPS could promote the production of Th1 cytokines but had no significant effect on IL17 expression. CONCLUSION: Antigen specific Th17 cells are distinct from antigen specific Th1 and Th2 cells. TGFβ, IL6 and IL23 are the factors responsible for promoting the differentiation and development of Th17 subset, whereas IFNγ and IL4 have inhibitory effects. 　　[Keywords]Th17 cells; T cell subset; flow cytometry; cytokine 　　按照CD4+T细胞分化和功能特征可以将其分为Th1和Th2细胞[1]。Th1型细胞通过分泌干扰素γ(IFNγ)等细胞因子介导细胞免疫; Th2型细胞通过分泌IL4等细胞因子介导体液免疫[2, 3]。Th1型免疫反应失调, 出现组织损坏和慢性炎症, 而Th2型免疫反应失调, 则导致过敏和哮喘疾病的发生[3]。近年来, 在类风湿关节炎、 实验性自身免疫性脑脊髓炎等自身免疫病和过敏原特异性反应研究中发现一种特殊的辅助型T细胞。这群CD4+T细胞同传统Th1和Th2细胞具有明显不同的特征: 主要分泌IL17, 表达控制其分化的独特的转录因子——孤独受体（orphan nuclear receptor, RORγt）[4], 因此命名为Th17细胞。目前对于此细胞亚群的研究主要集中于其在疾病的发生发展中的作用, 而探讨影响抗原特异性Th17细胞的诱导和分化因素的报道较少。为此, 我们研究了体外培养体系中, 初始抗原特异性TCRTg CD4+T细胞, 经抗原刺