环孢素A抑制正常人外周血记忆CD4^+T细胞产生IL-22的实验研究

CsA inhibits the production of IL-22 by memory CD4^+ T cells of PBMCs from healthy donors

目的探讨环孢素A(CsA)是否能够抑制正常人CD4+T细胞产生白细胞介素22(IL-22)。方法使用密度梯度离心法制备正常人外周血单个核淋巴细胞(PBMCs),加入PMA+Inomycin刺激细胞后,做细胞内细胞因子染色,流式细胞仪检测正常人外周血PBMCs中产生IL-22的T细胞亚群,以及IL-17-IFN-γ-IL-22+CD4+T细胞亚群(Th22);PMA+Inomycin刺激正常人PBMCs,同时加或不加环孢素A(CsA)后,做细胞内细胞因子染色,流式细胞仪检测IL-22+CD4+T细胞和IFN-γ+CD4+T细胞的比例,以及IL-22+CD4+T细胞表达记忆表型分子CD45RO情况。结果正常人外周血中产生IL-22的T细胞以CD4+T细胞为主,且存在仅分泌IL-22,不分泌IL-17和IFN-γ的Th22亚群;CsA以剂量依赖方式抑制CD4+T细胞产生IL-22和IFN-γ;CsA抑制外周血PBMCs中记忆CD4+T细胞产生IL-22。结论 CsA能抑制正常人外周血中记忆CD4+T细胞产生IL-22。

Objective To evaluate whether cyclosporine A（CsA） could inhibit the production of IL-22 by CD4＋ T cells of PBMCs from healthy donors.Methods Peripheral blood mononuclear cells （PBMCs） were isolated from heparinized blood of healthy donors using ficoll-hypaque density gradient centrifugation,and were stimulated with PMA＋ionomycin.Intracellular staining and flow cytometric analysis was done to determine：IL-22 produced by CD4＋ T,CD8＋ T cells,IL-22＋CD4＋ T cells and Th22 cells of PBMCs.PBMCs were stimulated with PMA＋ionomycin without or with different concentration of cyclosporine A （CsA）.Intracellular staining and flow cytometric analysis was done to determine：the percentage of IL-22＋CD4＋ T and IFN-γ＋CD4＋ T cells,and CD45RO expression of IL-22＋CD4＋ T of PBMCs in the presence or absence of CsA.Results There is a subset of Th22 from PBMCs from healthy donors.CsA could inhibit the production of IL-22 and IFN-γ by CD4＋ T cells from healthy donors in a dose dependent manner.CsA inhibits the production of IL-22 by memory CD4＋ T cells from healthy donors.Conclusion CsA inhibits the production of IL-22 by memory CD4＋ T cells of PBMCs from healthy donors.