目的:探讨γδT17/Th17/Tc17细胞在H1N1重症感染小鼠肺脏中的分布及其与肺组织炎性损伤的关系.方法:通过滴鼻感染建立H1N1重症感染小鼠模型;采用流式细胞术检测小鼠肺组织中γδT17/Th17/Tc17细胞的比例和数目;采用ELISA和Luminex多因子试剂盒检测肺泡灌洗液(BALF)中IL-17A、IL-1β、IL-23的浓度和血清中IL-17A 的浓度.结果:①建立了H1N1重症感染小鼠模型;②感染后第3天小鼠肺脏中γδT细胞占总淋巴细胞的比例较对照组显著升高(P<0.01),而Th和Tc细胞的比例较对照组无明显差异;③感染后第1天肺脏中γδT17细胞占总γδT细胞的比例和数目较对照组显著升高(P<0.05);Th17和Tc17细胞占Th和Tc细胞的比例和数目较对照组也升高;其中γδT17细胞的数目显著高于Th17和Tc17细胞(P<0.05);④感染后小鼠BALF中IL-17A的浓度逐渐升高,与对照组相比有显著性差异(P<0.05);感染后第3天血清中IL-17A也显著升高(P<0.05);BALF中可能参与γδT17细胞活化的IL-1β和IL-23的浓度较对照组显著升高.结论:γδT17细胞有可能以γδTCR非依赖的作用方式活化,并通过释放IL-17A参与H1N1重症感染小鼠早期肺组织炎性损伤过程.%Objective:To investigate the distribution of γδT17,Th17 and Tc17 cells in the lung of mice severely infected by influenza A(H1N1)pdm09 virus and the relationship between these cells with lung immunopathalogical injury.Methods:Intranasal infection was used to establish mouse model of severe H1N1 infection.Flow cytometry assay was used to detect the proportion and number of γδT17 cells,Th17 cells and Tc17 cells in the lung.The concentrations of interleukin-17A(IL-17A),interleukin-1β(IL-1β)and interleukin-23(IL-23) in the bronchoalveolar lavage fluid and serum were assayed by enzyme-linked immunosorbent assay and Lu-minex assay.Results:①The model of mice severely infected by influenza A(H1N1)pdm09 virus was established successfully.②The ratio of γδT cells,but not CD4+T and CD8+T cells in total lymphocytes of the lung of infected mice significantly increased compared with uninfected control mice at the third day post infection(DPI)(P<0.01).③The proportion and number of γδT17 cells,Th17 cells and Tc17 cells in total γδT cells,Th cells and Tc cells in the lung of infected mice were significant higher than that in uninfected control mice at the first DPI,respectively.However,the absolute number of γδT17 cells was far more than Th17 and Tc17 cells(P<0.05);④The concentration of IL-17A in BALF increased significantly after infection(P<0.05),and the concentration of IL-17A in serum increased significantly at the third DPI(P<0.05).The concentrations of both IL-1β and IL-23 in BALF probably participating in the activation of γδT17 cells increased significantly after infection compared with uninfected control mice.Conclusion:The γδT17 cells could be activated and secreted IL-17A via γδTCR non-depended pathway and involved in inflammatory pathological injury of lung at the early stage of severe H1N1 infection.
展开▼