摘要
目的:研究猪布鲁菌S2株诱导巨噬细胞(Mφ)的凋亡、活化及对T淋巴细胞的激活作用。方法:用猪布鲁菌S2株体外感染Mφ,并以大肠埃希菌作为对照,采用瑞氏-姬姆萨染色观察细胞形态学变化;流式细胞仪测定感染后24小时Mφ的凋亡率;ELISA法检测感染后不同时间点Mφ培养上清中IL-12和TNF-α及感染后Mφ与T细胞共培养上清中IFN-γ的含量的变化。结果:瑞氏-姬姆萨染色显示布鲁菌S2株感染Mφ后1小时体积明显增大,突起增多,胞浆内可见大量布鲁菌;6小时细胞内细菌减少,但胞膜仍完整;至12小时细胞核固缩,胞膜失去完整性。流式细胞仪检测结果显示,感染后24小时Mφ的凋亡率均明显高于正常对照组及大肠埃希菌感染组(P<0.05)。ELISA结果显示S2株感染Mφ12、24、48小时的上清中IL-12和TNF-α含量均明显高于正常对照组(P<0.01),S2株感染后Mφ与T细胞共培养24、48、72小时产生的IFN-γ量高于正常对照组(P<0.05),但低于大肠埃希菌感染组(P<0.01)。结论:S2株可以引起Mφ的凋亡,同时可诱导Mφ活化,产生IL-12和TNF-α;布鲁菌感染后的Mφ可诱导T细胞活化,产生IFN-γ免疫应答。
Objective:To study the apoptosis and activation of macrophages and their abilities to initiate immune actvation induced by B.suis S2.Methods: The macrophages infected by B.suis S2 were compared with Escherichia coli in vitro.The cell morphology changes were observed by Wrights-Giemsa stain and the cell apoptosis rates at 24 h post-infected were determined by Flow Cytometry.The IL-12 and TNF-α levels secreted by macrophages and the IFN-γ levels in the supernatants of T cells co-cultured with macrophages at different time points were detected by ELISA.Results: After 1 h infected by B.suis S2,the volume of macrophages was significantly expanded.The protuberances were increased and there were a large number of B.suis S2 in the cytoplasm of macrophages;the number of B.suis S2 decreased and the cell membranes remained intact after 6 h;the macrophages appeared karyopyknosis and the membranes were not intact after 12 h.The apoptosis rates of macrophages at 24 h post-infected by B.suis S2 were significantly higher than those of control group and Escherichia coli group(P〈0.05).The IL-12 and TNF-α levels of macrophages infected by B.suis S2 were significantly higher than those of the control group at 12,24 and 48 h(P〈0.01).The IFN-γ levels released from T cells co-cultured with macrophages at 24,48 and 72 h were also significantly higher than those of the control group(P〈0.05),but lower than those of Escherichia coli group(P〈0.01).Conclusion: B.suis S2 can induce Mφ apoptosis,activation and secretion of IL-12 and TNF-α;Mφ infected by B.suis S2 can induce IFN-γ production of T cell.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2013年第8期800-804,共5页
Chinese Journal of Immunology
基金
河北省自然基金资助项目(C2010000243)