常用免疫抑制剂对Tfr细胞和Breg细胞的作用研究
Effect of common immunosuppressants on Tfr cells and Breg cells
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摘要:目的 探讨常用免疫抑制剂对滤泡调节性T细胞(Tfr细胞)和调节性B细胞(Breg细胞)的作用。方法 收集6例健康志愿者的外周血(每例5 mL), 采用人淋巴细胞分离液分离出淋巴细胞进行培养, 将细胞分为对照组(未加入免疫抑制剂)、环孢素组(250 ng/mL)、他克莫司组(10 ng/mL)和西罗莫司组(10 ng/mL), 分别用相应免疫抑制剂刺激48 h。用流式细胞仪检测各组淋巴细胞中Breg细胞和Tfr细胞的比例; 采用酶联免疫吸附试验(ELISA)检测各组上清液中白细胞介素(IL)-21的含量。结果 与对照组比较, 环孢素组、他克莫司组和西罗莫司组淋巴细胞中Breg细胞的比例均升高, 差异均有统计学意义(均为P < 0.05);环孢素组和他克莫司组淋巴细胞中Tfr细胞比例均升高, 差异均有统计学意义(均为P < 0.05);西罗莫司组淋巴细胞中Tfr细胞比例下降, 差异亦有统计学意义(P < 0.05)。ELISA检测结果显示, 与对照组比较, 环孢素组和他克莫司组上清液中IL-21含量均下降, 差异均有统计学意义(均为P < 0.05);西罗莫司对IL-21分泌的影响不大, 差异无统计学意义(P > 0.05)。结论 环孢素和他克莫司具有抑制B细胞免疫反应的作用。Abstract:Objective To investigate the effect of common immunosuppressants on the follicular regulatory T cells (Tfr cells) and regulatory B cells (Breg cells).Methods Peripheral blood samples were collected from 6 healthy volunteers (5 mL per case).The lymphocytes were isolated by human lymphocyte isolation solution and cultured.The cells were divided into the control group (without supplement of immunosuppressants), ciclosporin group (250 ng/mL), tacrolimus group (10 ng/mL) and sirolimus group (10 ng/mL).The cells were stimulated with corresponding immunosuppressants for 48 h, respectively.The proportion of Breg and Tfr cells in the lymphocytes in each group was detected by flow cytometry.The content of interleukin (IL)-21 in the supernatants of each group was detected by enzyme-linked immunosorbent assay (ELISA).Results Compared with the control group, the proportion of Breg cells in the lymphocytes in the ciclosporin, tacrolimus and sirolimus groups was all significantly increased (all P < 0.05).In the ciclosporin group and tacrolimus group, the proportion of Tfr cells in the lymphocytes was both significantly elevated (both P < 0.05), whereas the proportion of Tfr cells in the lymphocytes in the sirolimus group was significantly decreased (P < 0.05).ELISA demonstrated that compared with the control group, the IL-21 content in the supernatants in the ciclosporin and tacrolimus groups was both significantly decreased (both P < 0.05).Use of sirolimus exerted no significant effect upon the secretion of IL-21 (P > 0.05).Conclusions Ciclosporin and tacrolimus can inhibit the immune response of B cells.
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