Induction of T regulatory cells by cytotoxic T-lymphocyte antigen-2alpha on corneal endothelial cells
Induction of T regulatory cells by cytotoxic T-lymphocyte antigen-2alpha on corneal endothelial cells
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细胞毒性 T 淋巴细胞抗原 2α 对角膜内皮细胞诱导 T 调节细胞
DOI：
10.1167/iovs.10-6322
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Mochizuki M
中科院分区：
文献类型：
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作者：
Sugita S;Yamada Y;Horie S;Nakamura O;Ishidoh K;Yamamoto Y;Yamaguchi S;Mochizuki M
目的：为了确定小鼠角膜内皮（CE）细胞是否可以促进调节性T细胞（Treg）的体外生成。为了通过CE细胞系体外诱导Treg细胞，将暴露于CE细胞的T细胞用作Treg细胞。收获在抗小鼠CD 3抗体存在下暴露于CE细胞的T细胞，并在体外添加至靶旁观者T细胞。通过[3 H]-胸苷掺入评估T细胞活化的增殖。通过流式细胞术评估Treg细胞上的⑶ 25或Foxp 3的表达。采用流式细胞术、RT-PCR、免疫组化和原位杂交检测CE细胞上细胞毒性T淋巴细胞抗原-2 α（CTLA-2α）的表达。使用抗CTLA-2α中和抗体、CTLA-2α siRNA或组织蛋白酶L前体阻断蛋白来消除CE抑制功能。培养的CE细胞在其表面产生CTLA-2α，从而使旁观者CD 4 + T细胞能够通过TGFβ促进转化为Treg细胞。CE诱导的Treg细胞通过高表达CD 25 high和Foxp 3而具有免疫抑制能力。当使用mRNA下调（siRNA转染）、中和抗体或阻断蛋白来阻断CE细胞上CTLA-2α的表达时，CE诱导的Treg细胞不能获得Treg功能。这些发现表明细胞表面CTLA-2α有助于旁观者T细胞的CE依赖性抑制。因此，眼部驻留组织暴露的T细胞可以被诱导成为外周微环境内的调节因子。
Purpose.: To determine whether murine corneal endothelial (CE) cells can promote the generation of T regulatory (Treg) cells in vitro.Methods.: To induce Treg cells in vitro by CE cell lines, T cells exposed to CE cells were used as Treg cells. T cells exposed to CE cells in the presence of anti–mouse CD3 antibody were harvested and added to target bystander T cells in vitro. T-cell activation was assessed for proliferation by [3 H]-thymidine incorporation. Expression of CD25 or Foxp3 on Treg cells was evaluated by flow cytometry. Expression of cytotoxic T-lymphocyte antigen-2 alpha (CTLA-2α) on CE cells was evaluated by flow cytometry, RT-PCR, immunohistochemistry, or in situ hybridization. Anti–CTLA-2α neutralizing antibodies, CTLA-2α siRNA, or pro-cathepsin L blocking proteins were used to abolish the CE-inhibitory function.Results.: Cultured CE cells produced CTLA-2α on their surfaces, thereby enabling bystander CD4+ T cells to be converted to Treg cells by TGFβ promotion. CE-induced Treg cells had immunosuppressive capacities by highly expressing CD25 high and Foxp3. When mRNA downregulation (siRNA transfection), neutralizing antibodies, or blocking proteins were used to block CTLA-2α expression on CE cells, CE-induced Treg cells failed to acquire Treg function.Conclusions.: These findings indicate that cell surface CTLA-2α contributes to the CE-dependent suppression of bystander T cells. Thus, ocular resident tissue-exposed T cells can be induced to become regulators within the peripheral microenvironment.