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目的 研究调节性T细胞(Treg)对变态反应性肺泡炎发生发展过程中CTLA-4分子和抑制性细胞因子水平的影响.方法 腹腔注射抗CD25单克隆抗体建立小鼠Treg消除模型,通过气管灌注1-3-β葡聚糖建立小鼠变态反应性肺泡炎模型,采用流式细胞术检测小鼠肺门淋巴结和脾细胞CD4+T细胞表面CTLA-4的表达水平;酶联免疫吸附法检测小鼠肺泡灌洗液中抑制性细胞因子的分泌水平.结果 与葡聚糖模型组小鼠比较,Treg消除组小鼠肺泡灌洗液中CD4+CD25+Treg百分比[(1.72±0.19)%]明显降低(P<0.05);葡聚糖灌注后早期,Treg消除组小鼠肺门淋巴结和脾细胞中CD4+CTLA-4+T细胞的比例[分别为(0.73±0.26)%、(2.69±0.17)%]较葡聚糖模型组小鼠[分别为(3.59±0.59)%、(15.80±3.54)%]明显降低,差异有统计学意义(P<0.05);葡聚糖灌注后晚期,Treg消除组小鼠肺泡灌洗液中TGF-β和IL-10的分泌水平[分别为(714.5262±57.0194)pg/ml、(1054.8323±14.0614)pg/ml],与葡聚糖模型组相比[分别为(1075.6619±103.9167)pg/ml、(1383.8642±71.1598)pg/ml],明显降低,差异有统计学意义(P<0.05).结论 Treg调控变态反应性肺泡炎的发生发展主要是通过影响早期辅助性T细胞表面CTLA-4的表达及影响晚期抑制性细胞因子TGF-β和IL-10的分泌水平.“,”ObjectiveThe aim of the study was to demonstrate the influence of regulatory T cell(Treg)on the CTLA-4 and inhibitory cytokines during the development of hypersensitivity pneumonitis.Methods Injection of anti-CD25 mAb was used to setup the Treg depletion mice model.Intratrachea instillation of 1,3-β-glucan was used to setup the hypersensitivity pneumonitis mice model.Flow cytometry was used to assay the CTLA-4 expression on CD4+T cell in mice hilar lymph node(HLN)and spleen.Enzyme-linked immunosorbent assay(ELISA)was used to examine the secretions of inhibitory cytokines in mice bronchoalveolar lavage fluid(BALF).Results The percentage of CD4+CD25+Treg in the Treg depletion group mice(1.72±0.19)% was significantly lower than that of the glucan group mice(P<0.05).According to flow cytometry assay,in the early stage after glucan instillation,the percentage of CD4+CTLA-4+T cells in mice HLN of the Treg depletion group (0.73±0.26)% was significantly reduced compared with that in the glucan group(3.59±0.59)% (P<0.05).Similarly,he percentage of CD4+CTLA-4+T cells in spleen of the Treg depletion group(2.69±0.17)% was significantly reduced compared with that in the glucan group(15.80±3.54)% (P<0.05).According to ELISA results,in the late stage after glucan instillation,the secretions of TGF-β(714.5262±57.0194)pg/ml and IL-10(1054.8323±14.0614)pg/ml in mice BALF of the Treg depletion group was distinctly decreased compared with that of the glucan group(1075.6619±103.9167)pg/ml and(1383.8642±71.1598) pg/ml (P<0.05).Conclusions Treg can regulate the development of hypersensitivity pneumonitis mainly through affecting the CTLA-4 expression on T-helper cells during the early stage and the secretions of inhibitory cytokines TGF-β and IL-10 during the late stage.