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目的 探讨γδT细胞在哮喘的免疫应答模式 ,认识γδT细胞亚群在哮喘发病机制中的作用。方法 Wistar大鼠 2 0只 ,随机分为健康对照组与哮喘组 (用鸡卵清蛋白致敏和刺激大鼠 ,制作哮喘模型 ) ,每组 10只。收集外周血单个核细胞 (PBMC)和支气管肺泡灌洗液 (BALF) ,用补体攻击法结合洗淘法选择性培养扩增γδT细胞 ,并用流式细胞术鉴定培养体系中的γδT细胞及其纯度 ,用原位杂交法测γδT细胞白细胞介素 (IL) 4mRNA和干扰素 (IFN)γmRNA的表达 ,用ELISA检测培养上清液中IL 4和IFN γ的浓度。结果 哮喘组大鼠PBMC和BALF中 ,γδT细胞培养上清液中IL 4浓度显著高于健康对照组 (P <0 0 1) ,IFN γ浓度低于健康对照组 (P <0 0 1) ;γδT细胞IL 4mRNA表达阳性率高于健康对照组 (P <0 0 1) ,IFN γmRNA表达阳性率低于健康对照组 (P <0 0 1)。结论γδT细胞或者γδT细胞亚群存在Th1/Th2模式 ,在大鼠哮喘模型呈Th2优势应答 ,γδT细胞参与了哮喘的发病过程。
Objective To investigate the immune response of γδT cells to asthma and to understand the role of γδT cell subsets in the pathogenesis of asthma. Methods Twenty male Wistar rats were randomly divided into healthy control group and asthma group (sensitized and stimulated with chicken ovalbumin to make asthma model), with 10 rats in each group. Peripheral blood mononuclear cells (PBMCs) and bronchoalveolar lavage fluid (BALF) were collected, and γδT cells were selectively expanded by complement challenge and wash-washing method. The γδT cells in culture system were identified by flow cytometry and their purity , The expression of interleukin (IL) 4 mRNA and interferon (IFN) γ mRNA in γδT cells was detected by in situ hybridization, and the concentrations of IL 4 and IFN γ in the culture supernatants were detected by ELISA. Results The concentration of IL - 4 in the culture supernatant of γδT cells in PBMC and BALF of asthmatic rats was significantly higher than that of healthy controls (P <0.01), and the concentration of IFNγ was lower than that of healthy controls (P <0.01). The positive rate of IL 4 mRNA expression in γδT cells was higher than that in healthy controls (P <0.01), and the positive rate of IFNγmRNA expression was lower than that in healthy controls (P <0.01). Conclusions Th1 / Th2 mode exists in γδT cells or γδT cell subsets, Th2 predominant responders in rat asthma model, γδT cells are involved in the pathogenesis of asthma.