BCG-PPD、H37Rv-PPD诱导入巨噬细胞死亡及TNF-α、IL-1β和IL-10的表达差异  

Different cell death of THP-1 induced by virulent/attenuated purfied protein derivatives tuberculin and the different expression of TNF-α,IL-1β and IL-10 in Mycobacterium tuberculosis

作　　者：史会连[1] 路蝉伊[1] 虞胜镭[1] 张文宏[1] 翁心华[1] 陈澍[1] 

机构地区：[1]上海复旦大学附属华山医院感染科,200040

出　　处：《中华微生物学和免疫学杂志》2010年第9期834-837,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家“十一·五”重大传染病防治专项课题（2008ZX10003-011）

摘　　要：目的 本实验对不同毒力结核杆菌的衍生蛋白（PPD）对人巨噬细胞（THP-1）的影响及其与TNF-αt、IL-1 β及IL-10的差异性进行研究.方法 用H37Rv-PPD和BCG-PPD分别在3 h、8 h、15 h及24 h四个时间点刺激分化成熟的THP-1细胞,再应用Hochest染色法,荧光镜下观察细胞的转归差异（凋亡及坏死情况）,同时取上清用ELISA法测TNF-α、IL-1β及IL-10的浓度.结果 BCG-PPD刺激下细胞核以椭圆凋亡小体多见,而H37Rv-PPD刺激下细胞核则多呈坏死状,以坏死多见;BCG-PPD刺激下上清中TNF-α及IL-10的表达量低于H37Rv-PPD刺激组,但BCG-PPD刺激下IL-1β的表达量却高于后者.结论 提示高毒力菌株衍生蛋白（H37Rv-PPD）引起THP-1坏死的原因可能与TNF-α的过度表达有关,而凋亡少见可能与IL-10抑制凋亡作用有关,而低毒力菌株衍生蛋白诱导凋亡与IL-1β有关.可能菌株毒力差异就存在于菌株的蛋白成分之中,且与上述几种细胞因子密切相关.Objective To study the different response in macrophages treated with different agoβ and IL-10 in Mycobacterium tuberculosisnists（H37Rv-PPD and BCG-PPD）related with Mycobacterium tuberculosis and the relationship with TNF-αt,IL-1β and IL-10.Methods Using H37Rv-PPD and BCG-PPD to stimulate THP-1 cell for 3h,8h,15h,24h respectively.Cells were ananlyzed by Hochest staining under fluorescence microscopy to assay cell death（apoptosis and necrosis）.At each stimulating time,TNF-α,IL-1β and IL-10 were examined by ELISA.Results Under fluorescence microscopy,it could easily see oval apoptotic bodies of THP-1 stimulated by BCG-PPD.However ,the nucleus were often isolated and necrosis-like when cells were stimulated by H37Rv-PPD.In a word ,BCG-PPD tend to induce THP-1 cells to apoptosis,but H37Rv-PPD inclined to induce cells to undergo necrosis.In supernatant of cells stimulated by BCG-PPD,the expression of TNF-αand IL-10 were lower than the cells stimulated by H37Rv-PPD,but the expression of IL-1β was higher than the latter.Conclusion It indicated that the necrosis of cells stimulated by H37Rv-PPD was asossiated with the excessive expression of TNF-α and IL-10,and the apoptosis of cells induced by BCG-PPD was IL-1β related.Perhaps the mechanism of differences in virulence exist in protein of strain,and associated with cytokines IL-1β,TNF-α and IL-10.

关 键 词：结核分枝杆菌 THP-1 细胞死亡 

分 类 号：R392.1[医药卫生—免疫学]