评估人T淋巴细胞亚群对结核杆菌脂质抗原的免疫反应  

Evaluation of immune responses of human T lymphocyte subsets to Mycobacterium tuberculosis lipid antigens

作　　者：金亮[1] 张茜[1] 李柏青[1] 沙泉[1] 

机构地区：[1]安徽医科大学免疫学教研室及过敏与免疫研究中心,合肥230032

出　　处：《中国免疫学杂志》2016年第2期159-164,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金(No.81273245);安徽省国际科技合作计划(No.11030603027)

摘　　要：目的:观察结核杆菌脂质的一些脂类抗原对人外周血中淋巴细胞活化和增殖的作用,进而探讨脂类抗原在结核感染中是否存在特异性的免疫作用。方法:取健康人外周血单个核细胞(PBMC),分离诱导出非成熟树突状细胞(im DC)后分别加入结核杆菌脂质全脂质(TLIP)、丙酮可溶性脂质(ASLIP)、纯硫脂(PSLIP)、脂阿拉伯糖(LAM)、脂甘露聚糖(LM)、同时设置非脂类抗原全菌裂解物(WCL)、培养分泌蛋白(CFP)、耐热抗原(Mtb-HAg)、脂多糖(LPS)和空白对照组。用CFSE标记自体淋巴细胞后,与被脂质抗原刺激后的DC共培养。之后,用流式细胞仪(FCM)检测T淋巴细胞亚群(CD4+、CD8+、NKT和γδT)的增殖和分泌细胞因子(IFN-γ、TNF-α和IL-4)的情况。结果:相较于空白组,所有脂质都能促进NKT细胞和CD8+T细胞增殖(P<0.05)。相较于空白组ASLIP,LAM和LM可以促进非增殖的CD4+T细胞分泌IL-4,或者促进增殖的CD4+T细胞分泌IFN-γ(P<0.05)。相较于空白对照组,所有脂质抗原(除了LM)都能促进增殖的γδT和CD8+T细胞分泌IFN-γ和TNF-α,而LM能抑制增殖的CD8+T细胞分泌TNF-α。结论:结核杆菌脂质抗原能激活PBMC的特异性免疫反应,特别是CD1限制性T细胞的应答。Objective：Some antigens of M. tb to culture with peripheral blood mononuclear cells （PBMC） for assaying their proliferation and activation, so as to signify whether lipid antigens of M. tb have specific immune responses in host against M. tb infection or not. Methods： We treated PBMC with several lipid antigens of M. tb to explore the ability of these antigens to activate immunity in healthy individuals. We measured and analyzed cell proliferation by labeling cells with carboxyfluorescein succinimidyl amino ester （CFSE） and subjecting them to flow cytometry （FCM）. The production of IFN-y, TNF-a and IL-4 by T cell subsets （NKT, CD4＋, CD8＋, and yδT） from healthy donors was analyzed by FCM after stimulation with autologous immature dendritic cells pre-cultured with M. tb lipid antigens. The tested M. tb lipid antigens were the total lipid （TLIP）, Acetone-Soluble Lipids （ASLIP）, Purified Sulfolipid （ PSLIP）, Lipoarabinomannan （ LAM ） and Lipomannan （ LM ） levels. Medium free of lipid antigens （ WCL, CFP, LPS, Mtb-HAg and blank） was used as a control. Results： We found the proportion of proliferative NKT and CD8 ＋T cells significantly increased in all lipid groups （P〈0. 05）. ASLIP, LAM and LM promoted non-proliferative CD4＋T ceils to secrete IL-4 and proliferative ones to secrete IFN-y （P〈0. 05 ）. All lipid antigens promoted both proliferative yδT cells and CD8＋T cells to secrete IFN-y/and TNF-a, but the proportion of TNF-a-secreting cells in these populations decreased in the LM group （P〈0. 05）. Conclusion： Lipid antigens may affect the CD1restricted T cells of the host to fight M. tb infection.

关 键 词：结核杆菌脂质抗原 细胞因子 T淋巴细胞亚群 增殖反应 

分 类 号：R329.4[医药卫生—人体解剖和组织胚胎学]