结核杆菌抗原对人γδT细胞体外增殖影响  

Effects of mycobacterium tuberculosis antigen on proliferation of human γδT cellsin vitro

作　　者：佟伟[1] 李宏伟[1] 潘兴瑜[2] 

机构地区：[1]辽宁医学院免疫与病原生物学教研室,锦州121001 [2]中国医科大学免疫学教研室

出　　处：《中国公共卫生》2008年第6期670-671,共2页Chinese Journal of Public Health

摘　　要：目的观察结核杆菌抗原(Mtb-Ag)对人γδT细胞体外增殖的影响,以解决医学研究中γδT细胞来源不足问题。方法应用酸性改良罗氏培养基分离培养结核杆菌及丙酮酸钠液体培养基进行扩增,通过85℃水浴灭活,超声波细胞破碎制备Mtb-Ag。在体外以不同的剂量刺激人外周血单个核细胞(PBMC),采用四甲基偶氮噻唑蓝(MTT)法检测增殖程度,应用流式细胞仪检测γδT细胞增殖的百分率。结果在一定范围内随着刺激剂量的增加PBMC增殖亦增加。流式细胞仪分析增殖结果显示,7μg/ml刺激组γδT细胞的增殖率最高(约为49.08%)。但当刺激剂量增加至9μg/ml时,γδT细胞增殖比率开始降低(约为32.62%),而αβT细胞增殖比率增加明显(约为36.76%)。结论Mtb-Ag在合适刺激剂量时,可对人γδT细胞发挥优势扩增;当剂量过大时则对αβT细胞发挥优势扩增。Objective To observe the effects of mycobacterium tuberculosis antigen on the proliferation of human γδT cells in vitro and resolve the problem of γδT cells shortage. Methods Acid L - G medium was used to isolate mycobacterium tuberculosis（Mtb） and sodium pyruvate medium was employed to proliferate the bacteria. Soluble extracts Mtb-Ag was obtained by heat - treating at 85℃ and sequentially disrupted by prolonged sonication. Mtb - Ag was used to stimulate peripheral blood mononuclear cells at different doses in vitro. The proliferation of PBMC was detected by MTT chromatometry method during culture. The percentage of γδT cells was measured by flow cytometry. Results The capability of Mtb antigens to stimulate PBMC proliferation varied with the dose. PBMC proliferation was dose - dependent from Mtb antigens in given range. It was demonstrated by flow cytometry that 7μg/ml dose Mtb - Ag group could stimulate γδT cells remarkably to the highest level（approximately 49.08 % ）. But when the stimulation dose increased to 9 vg/ml, the percentage of γδT cells started to decrease（approximately 32.62 % ） while T cell ratio increased markedly（approximately 36.76 % ）. Conclusion In vitro, the component of Mtb - Ag can effectively stimulate the proliferation of human γδT cells. The results indicates that Mtb - Ag can dominantly induce γδT cells proliferation at proper dose and activate αβT cells only at higher dose.

关 键 词：结核杆菌抗原 ΓΔT细胞 增殖 

分 类 号：R378.91[医药卫生—病原生物学]