Overexpression of T-bet gene regulates murine autoimmune arthritis
Overexpression of T-bet gene regulates murine autoimmune arthritis
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DOI：
10.1002/art.33335
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发表时间：
2012-01-01
影响因子：
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通讯作者：
Sumida, Takayuki
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文献类型：
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作者：
Kondo, Yuya;Iizuka, Mana;Sumida, Takayuki
目的探讨T-bet在胶原诱导性关节炎（CIA）发病机制中的作用。方法.产生在CD 2启动子控制下的T-bet转基因（Tg）小鼠。在T-bet-Tg小鼠和野生型C57 BL/6（B6）小鼠中诱导CIA。通过实时聚合酶链反应分析II型胶原（CII）反应性T-bet和视黄酸受体相关孤儿核受体γ t（ROR γ t）信使RNA表达水平。采用来自B6和T-bet-Tg小鼠的CD 4 + T细胞以及来自患有CII的B6和T-bet-Tg小鼠的CD 11 c+脾树突状细胞（DC）进行交叉实验，并通过酶联免疫吸附测定法测定上清液中的白细胞介素-17（IL-17）和干扰素-γ（IFN-γ）。培养来自B6、T-bet-Tg或T-bet-Tg/IFN γ（-/-）小鼠的CD 4 + T细胞用于Th 17细胞分化，然后通过荧光激活细胞分选分析产生IFN α和IL-17的细胞的比例。结果与B6小鼠不同，T-bet-Tg小鼠没有发生CIA。T-bet-Tg小鼠显示Tbx 21过表达和CII反应性T细胞中Rorc下调。用CD 4 + T细胞和脾DC进行的交叉实验显示，即使与来自B6小鼠的DC共培养，T-bet-Tg小鼠中CII反应性CD 4 + T细胞的IL-17产生也显著减少，表明产生IL-17的CD 4 + T细胞功能障碍。在T-bet-Tg小鼠和T-bet-Tg/IFN γ（-/-）小鼠中均观察到在有利于Th 17细胞分化的体外条件下抑制Th 17细胞分化。结论T细胞中T-bet的过表达抑制了自身免疫性关节炎的发展。关节炎的调节机制可能涉及通过IFN γ非依赖性途径过表达T-bet导致CII反应性Th 17细胞分化障碍。
Objective To clarify the role of T-bet in the pathogenesis of collagen-induced arthritis (CIA). Methods. T-bet-transgenic (Tg) mice under the control of the CD2 promoter were generated. CIA was induced in T-bet-Tg mice and wild-type C57BL/6 (B6) mice. Levels of type II collagen (CII)-reactive T-bet and retinoic acid receptor-related orphan nuclear receptor gamma t (ROR gamma t) messenger RNA expression were analyzed by real-time polymerase chain reaction. Criss-cross experiments using CD4+ T cells from B6 and T-bet-Tg mice, as well as CD11c+ splenic dendritic cells (DCs) from B6 and T-bet-Tg mice with CII were performed, and interleukin-17 (IL-17) and interferon-gamma (IFN gamma) in the supernatants were measured by enzyme-linked immunosorbent assay. CD4+ T cells from B6, T-bet-Tg, or T-bet-Tg/IFN gamma(-/-) mice were cultured for Th17 cell differentiation, then the proportions of cells producing IFN alpha and IL-17 were analyzed by fluorescence-activated cell sorting. Results. Unlike the B6 mice, the T-bet-Tg mice did not develop CIA. T-bet-Tg mice showed overexpression of Tbx21 and down-regulation of Rorc in CII-reactive T cells. Criss-cross experiments with CD4+ T cells and splenic DCs showed a significant reduction in IL-17 production by CII-reactive CD4+ T cells in T-bet-Tg mice, even upon coculture with DCs from B6 mice, indicating dysfunction of IL-17-producing CD4+ T cells. Inhibition of Th17 cell differentiation under an in vitro condition favoring Th17 cell differentiation was observed in both T-bet-Tg mice and T-bet-Tg/IFN gamma(-/-) mice. Conclusion. Overexpression of T-bet in T cells suppressed the development of autoimmune arthritis. The regulatory mechanism of arthritis might involve dysfunction of CII-reactive Th17 cell differentiation by overexpression of T-bet via IFN gamma-independent pathways.