T淋巴细胞在HPV感染小鼠生殖道黏膜上皮细胞免疫反应中的作用

Effect of T lymphocyte on immune response of epithelial cells of genital tract mucosa of rats with HPV infection

目的 探讨T淋巴细胞在人乳头瘤病毒(HPV)感染小鼠模型生殖道黏膜上皮细胞免疫反应中的作用。方法 采用随机数字法将C57BL/6小鼠分为对照组、空载组、模型A组、模型B组,各12只,构建HPV16感染小鼠模型,进行小鼠生殖道分泌物、外周血、黏膜组织中炎性细胞分类计数;观察生殖道黏膜上皮病理状态;流式细胞仪检测外周血T淋巴细胞激活水平;实时荧光定量聚合酶链反应(RT-PCR)检测黏膜组织CD_4~+T淋巴细胞转录因子T盒子21转录因子(T-bet)、GATA结合蛋白3(GATA3)、维A酸孤儿核受体γt(ROR-γt)、叉头样转录因子P3(FOXP3),以及炎性因子白细胞介素(IL)-6、IL-10、肿瘤坏死因子-α(TNF-α)、转化生长因子-β(TGF-β)、干扰素-γ(IFN-γ)的mRNA转录水平、蛋白相对表达水平。结果 与对照组、空载组比较,模型A组、模型B组生殖道分泌物、外周血、黏膜组织中的炎性细胞数量高,外周血中CD_(3)^(+)T淋巴细胞亚群激活水平降低,CD_(4)^(+)、CD_(8)^(+)T淋巴细胞亚群激活水平升高,CD_(4)^(+)/CD_(8)^(+)比值降低,黏膜组织中CD_(4)^(+)T细胞亚群转录因子T-bet、GATA3、RORγt、FOXP3的mRNA转录水平高,以及T细胞亚群特异细胞因子IFN-γ、IL-4、IL-17A、FOXP3表达水平高,黏膜组织中炎性因子IL-6、TNF-α、IL-10、TGF-β、IFN-γ的mRNA转录水平、蛋白相对表达水平高,且模型B组高于模型A组(P<0.05)。结论 T淋巴细胞亚群通过不同的激活水平参与机体对HPV感染的免疫反应,其激活水平随HPV拷贝数而改变;其中CD_(4)^(+)在免疫应答的调节和效应中起着重要作用,通过调节炎性细胞因子的分泌达到抗病毒的作用。

OBJECTIVE To explore the effect of T lymphocyte on immune response of epithelial cells of genital tract mucosa of rat models with human papillomavirus(HPV)infection.METHODS Totally C57BL/6 rats were ran-domly divided into the control group,the no-load group,the model A group and the model B group,with 12 rats in each group.The rat models of HPV16 infection were established.The differential counting of inflammatory cells in genital tract secretions,peripheral blood and mucosal tissues was carried out for the rat models.The epithelial pathological state of genital tract mucosa was observed,the activation level of peripheral blood T lymphocyte was detected by flow cytometry;the mRNA transcription levels and relative expression levels of T box 21 transcription factor(T-bet),GATA binding protein 3(GATA3),retinoic acid orphan nuclear receptorγt(ROR-γt)and fork-head box P3(FOXP3)of CD_(4)^(+)T lymphocyte in mucosa tissues as well as inflammatory factors interleukin(IL)-6,IL-10,tumor necrosis factor-α(TNF-α),transforming growth factor-β(TGF-β)and interferon-γ(IFN-γ)were de-tected by means of real-time fluorescent quantitative polymerase chain reaction(RT-PCR).RESULTS The levels of inflammatory cells in genital tract secretions,peripheral blood and mucosa tissues were higher in the model A group and the model B group than in the control group and the no-load group;the activation levels of CD_(3)^(+)T lym-phocyte subsets in peripheral blood were lower in the model A group and the model B group than in the control group and the no-load group;the activation levels of CD_(4)^(+) and CD_(8)^(+)T lymphocyte subsets were higher in the model A group and the model B group than in the control group and the no-load group;the ratio of CD_(4)^(+)to CD_(8)^(+) was reduced;the mRNA transcription levels of transcription factors T-bet,GATA3,RORγt and FOXP3 of CD_(4)^(+)T lymphocyte subsets in mucosa tissues were higher in the model A group and the model B group than in the con-trol group and the no-load group;the expression levels of specific cytokines of T lymphocyte subsets IFN-γ,IL-4,IL-17A and FOXP3 were higher in the model A group and the model B group than in the control group and the no-load group;the mRNA transcription levels and relative expression levels of inflammatory factors IL-6,TNF-a,IL-10,TGF-βand IFN-γin mucosa tissues were higher in the model A group and the model B group than in the control group and the no-load group,and the levels of above indexes of the model B group were higher than those of the model A group(P<0.05).CONCLUSION T lymphocyte subsets involve in the body's immune response to HPV infection by the activation levels,and the activation level changes with the copies of HPV;CD_(4)_(+)plays a sig-nificant role in the regulation of immune response and effect and achieves the antiviral effect by regulating the se-cretion of inflammatory cytokines.