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目的:研究抗CD40L单克隆抗体(抗CD40L mAb)对哮喘大鼠外周血和淋巴液中CD4+ CD25+调节性T细胞(Treg)数量及其功能的影响。方法:抗CD40L mAb处理血液和淋巴液中的单个核细胞(MNCs),流式细胞仪(FCM)检测MNCs中CD4+ CD25+ Foxp3+ T细胞的百分率,酶联免疫吸附试验(ELISA)检测细胞培养上清中IL-10和TGF-β1的含量。结果:体外培养72 h后,淋巴液来源MNCs中CD4+CD25+Foxp3+Treg的比例均明显高于血液来源的MNCs,末次激发后各组淋巴液和血液来源MNCs中CD4+ CD25+ Foxp3+ Treg比例随细胞收集时间距末次激发时间延长呈先升高后降低的趋势;哮喘组淋巴液和血液来源MNCs中CD4+ CD25+ Foxp3+ Treg比例均显著低于正常对照组和抗CD40L mAb组(P<0.05);末次激发后0、12 h收集的血液MNCs培养上清中抗CD40L mAb组TGF-β1的水平显著高于哮喘组和正常对照组(P<0.05);末次激发后24 h收集的淋巴液来源MNCs培养上清中抗CD40L mAb组IL-10的水平显著高于哮喘组和正常对照组(P<0.05)。结论:抗CD40L mAb促进CD4+ CD25+ Foxp3+ T细胞增殖,并在哮喘激发早期(0、12 h)促进血液来源CD4+ CD25+ Foxp3+ Treg细胞分泌TGF-β1,哮喘激发后期(24 h)促进淋巴液来源CD4+ CD25+ Foxp3+ Treg细胞分泌IL-10。
Objective: To investigate the effect of anti-CD40L monoclonal antibody (anti-CD40L mAb) on the number and function of CD4 + CD25 + regulatory T cells (Tregs) in peripheral blood and lymph of asthmatic rats. Methods: Mononuclear cells (MNCs) from blood and lymph were treated with anti-CD40L mAb. The percentage of CD4 + CD25 + Foxp3 + T cells in MNCs was detected by flow cytometry (FCM). The cell culture supernatants were detected by enzyme-linked immunosorbent assay In IL-10 and TGF-β1 content. Results: The percentage of CD4 + CD25 + Foxp3 + Tregs in MNCs from lymphocytes was significantly higher than that from MNCs from 72h after culture in vitro. The proportion of CD4 + CD25 + Foxp3 + Tregs in lymphocytes and blood of MNCs from the last challenge was CD40 + Foxp3 + Treg in lymphocytes and blood of MNCs in asthma group were significantly lower than those in normal control group and anti-CD40L mAb group (P <0.05); the last challenge The levels of TGF-β1 in the anti-CD40L mAb in the supernatant of MNCs collected at 0 and 12 h after the last challenge were significantly higher than those in the asthma group and the normal control group (P <0.05) The level of IL-10 in Qingzhong anti-CD40L mAb group was significantly higher than that in asthma group and normal control group (P <0.05). Anti-CD40L mAb can promote the proliferation of CD4 + CD25 + Foxp3 + T cells and promote the secretion of TGF-β1 from blood-derived CD4 + CD25 + Foxp3 + Treg cells in the early stage of asthma challenge (0,12 h) Foxp3 + Treg cells secrete IL-10.