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目的探讨环孢素A(CsA)对人扁桃体滤泡辅助性T(Tfh)细胞产生白细胞介素-21(IL-21)以及对CD40L和Bcl-6表达的影响。方法分离正常人外周血及扁桃体单个核细胞,利用PMA+Ionomycin刺激加入不同剂量的CsA,采用酶联免疫吸附试验(ELISA)和酶联免疫斑点试验(ELISPOT)检测IL-21和γ-干扰素(IFN-γ)的产生,利用流式细胞术检测扁桃体Tfh细胞中IL-21、IFN-γ、Bcl-6和CD40L的表达,同时检测Tfh细胞的表型特征。结果 CsA呈剂量依赖性抑制扁桃体Tfh细胞IL-21和IFN-γ的产生,并抑制Bcl-6和CD40L的表达。表型分析显示,CsA主要抑制记忆型Tfh细胞产生IL-21,即CD45RO+CCR7+/-。结论 CsA呈浓度依赖性地抑制人扁桃体Tfh细胞IL-21的产生并降低其Bcl-6和CD40L的表达,主要抑制记忆型Tfh细胞产生IL-21,为探讨环孢素A对Tfh细胞亚群免疫功能的影响提供了依据。
Objective To investigate the effect of CsA on the production of interleukin-21 (IL-21) and the expression of CD40L and Bcl-6 in human tonsil follicular T cells. Methods Peripheral blood and tonsillar mononuclear cells were isolated from healthy volunteers. Different doses of CsA were stimulated with PMA + Ionomycin. The levels of IL-21 and IFN-γ were detected by enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunosorbent assay (ELISPOT) The expression of IL-21, IFN-γ, Bcl-6 and CD40L in Tfh cells was detected by flow cytometry and the phenotypic characteristics of Tfh cells were also detected. Results CsA inhibited the production of IL-21 and IFN-γ in Tfh cells in a dose-dependent manner and inhibited the expression of Bcl-6 and CD40L. Phenotypic analysis showed that CsA mainly inhibited the production of IL-21 in memory-type Tfh cells, that is, CD45RO + CCR7 +/-. Conclusion CsA can inhibit the production of IL-21 and decrease the expression of Bcl-6 and CD40L in Tfh cells in a concentration-dependent manner, and mainly inhibit the production of IL-21 by memory Tfh cells. To investigate the effect of CsA on Tfh cell subsets The impact of immune function provides the basis.