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目的体外研究小鼠巨细胞病毒(murine cytomegalovirus,MCMV)感染细胞热休克蛋白70与细胞毒T细胞(CTL)活性相关性。方法体外培养小鼠胚肺细胞,在加入MCMV后16、24、48、72、120h分别收获细胞,用免疫组织化学方法检测HSP70表达情况;CTL活性检测组:体外培养小鼠胚肺细胞,设以下各组:正常细胞对照组,病毒感染组,病毒感染+放线菌素组,病毒感染+槲皮素组,病毒感染+HSP70抗体组,分别在以上时相选用51Cr释放试验检测CTL活性。结果MCMV感染诱导小鼠胚肺细胞表达HSP70,与正常细胞对照组比较,各时相HSP70表达强度差异均具有显著性,其中16h始升高,120h表达最强。病毒感染组:CTL活性升高,与其它各组比较,各时相差异均具有显著性意义,自16h始升高,120h达最高;放线菌素组、槲皮素组、HSP70抗体组,CTL活性均无显著性差异,自16h始升高,120h达最高,与正常细胞对照组比较,CTL活性均升高。结论MCMV感染细胞热休克蛋白70与CTL的活性具有一定相关性。
Objective To study the relationship between heat shock protein 70 (HSP70) and cytotoxic T lymphocyte (CTL) activity in murine cytomegalovirus (MCMV) infected cells in vitro. Methods Mouse embryo lung cells were cultured in vitro. Cells were harvested at 16,24,48,72 and 120 h after addition of MCMV, and the expression of HSP70 was detected by immunohistochemistry. CTL activity assay: cultured mouse embryo lung cells in vitro The following groups: normal cell control group, viral infection group, viral infection + actinomycin group, viral infection + quercetin group, viral infection + HSP70 antibody group, respectively, in the above time selected 51Cr release test CTL activity. Results The expression of HSP70 in mouse embryo lung cells was induced by MCMV infection. Compared with the normal control group, the expression intensity of HSP70 in each phase was significantly different. The expression of HSP70 increased at 16h and reached the peak at 120h. In viral infection group, the activity of CTL increased, compared with other groups, the differences in each phase were significant, starting at 16h and reaching the highest at 120h. Actinomycin, quercetin, HSP70 antibody group, CTL activity had no significant difference, starting from 16h, reached the highest at 120h, compared with the normal cell control group, CTL activity were increased. Conclusion The activity of heat shock protein 70 in MCMV infected cells has a certain correlation with CTL.