Activated γδ T Cells Promote the Activation of Uveitogenic T Cells and Exacerbate EAU Development
Activated γδ T Cells Promote the Activation of Uveitogenic T Cells and Exacerbate EAU Development
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DOI：
10.1167/iovs.10-6758
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发表时间：
2011-07-01
影响因子：
4.4
通讯作者：
Sun, Deming
中科院分区：
文献类型：
--
作者：
Nian, Hong;Shao, Hui;Sun, Deming
目的.目的探讨γ δ T细胞活化对葡萄膜炎性α β T细胞生成及实验性自身免疫性葡萄膜炎（EAU）发生的影响。γ δ T细胞分离自用致葡萄膜肽IRBP（1-20）免疫的B6小鼠，α β T细胞分离自免疫的TCR-δ（-/-）小鼠。通过将分离的γ δ T细胞在不含精氨酸的培养基中培养3至5天来制备静息γ δ T细胞，此时它们显示出CD 69表达的下调。通过将静息γ δ T细胞与抗γ δ TCR（GL 3）孵育2天来制备活化的γ δ T细胞。将应答α β T细胞与免疫抗原和抗原呈递细胞共培养。在加入或不加入纯化的γ δ T细胞的情况下，通过细胞内染色，然后在刺激后进行FACS分析来确定表达IL-17或IFN-γ的抗原特异性T细胞的数量。用ELISA法检测培养液中细胞因子的含量。高度富集的γ δ T细胞对EAU中的自身反应性α β T细胞发挥广泛不同的作用，这取决于γ δ T细胞的活化状态。而非活化的γ δ T细胞在体外和体内对感光细胞间类维生素A结合蛋白特异性α β T细胞的活化几乎没有影响，活化的γ δ T细胞促进葡萄膜原性T细胞的产生并加剧EAU的发展。γ δ T细胞的功能能力受到其活化状态的极大影响。活化的γ δ T细胞通过增加葡萄膜原性T细胞的活化而加重EAU。（Invest Ophthalmol维斯科学。2011;52：5920-5927）DOI：10.1167/iovs.10-6758
PURPOSE. To determine how the activation of gamma delta T cells affects the generation of uveitogenic alpha beta T cells and the development of experimental autoimmune uveitis (EAU).METHODS. gamma delta T cells were isolated from B6 mice immunized with the uveitogenic peptide IRBP(1-20) and alpha beta T cells from immunized TCR-delta(-/-) mice. Resting gamma delta T cells were prepared by culture of separated gamma delta T cells in cytokine-free medium for 3 to 5 days, when they showed downregulation of CD69 expression. Activated gamma delta T cells were prepared by incubating resting gamma delta T cells with anti-gamma delta TCR (GL3) for 2 days. Responder alpha beta T cells were cocultured with immunizing antigen and antigen-presenting cells. The numbers of antigen-specific T cells expressing IL-17 or IFN-gamma were determined by intracellular staining followed by FACS analysis after stimulation, with or without the addition of purified gamma delta T cells. The cytokines in the culture medium were measured by ELISA.RESULTS. Highly enriched gamma delta T cells exert widely different effects on autoreactive alpha beta T cells in EAU, depending on the activation status of the gamma delta T cells. Whereas nonactivated gamma delta T cells had little effect on the activation of interphotoreceptor retinoid-binding protein-specific alpha beta T cells in vitro and in vivo, activated gamma delta T cells promoted the generation of uveitogenic T cells and exacerbated the development of EAU.CONCLUSIONS. The functional ability of gamma delta T cells is greatly influenced by their activation status. Activated gamma delta T cells exacerbate EAU through increased activation of uveitogenic T cells. (Invest Ophthalmol Vis Sci. 2011;52:5920-5927) DOI: 10.1167/iovs.10-6758