不同亚群CD8^+ T细胞表型、功能分子及分化相关基因的表达  

Differences in phenotype,function and differentiation-related genes expression of different CD8^+ T cell subsets

作　　者：田永贵 张震[1,2,4] 尹婕 申春一 张毅 TIAN Yonggui;ZHANG Zhen;YIN Jie;SHEN Chunyi;ZHANG Yi(Biotherapy Center,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052;Henan Key Laboratory for Tumor Immunology and Biotherapy,Zhengzhou 450052;School of Life Sciences,Zhengzhou University,Zhengzhou 450001;Cancer Center,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052)

机构地区：[1]郑州大学第一附属医院生物细胞治疗中心,郑州450052 [2]河南省肿瘤免疫与生物治疗重点实验室,郑州450052 [3]郑州大学生命科学学院,郑州450001 [4]郑州大学第一附属医院肿瘤中心,郑州450052

出　　处：《郑州大学学报（医学版）》2019年第3期318-323,共6页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金资助项目(81771781;U1804281;81802857);国家重点研发计划重点专项基金资助项目(2018YFC1313400);河南省重大科技专项基金资助项目(161100311000)

摘　　要：目的:探讨不同亚群CD8^+ T细胞的表型、功能分子及分化相关基因表达的差异。方法:密度梯度离心法从健康人外周血中分离外周血单个核细胞,并用MACS细胞免疫磁珠分选获取CD8^+ T细胞。采用CD3、CD8、CD45RA、CCR7标记CD8^+ T细胞亚群,检测各亚群表面标志物(CD69、CD28、PD-1)及功能分子(IFN-γ、IL-2)的表达;采用qRT-PCR检测CD8+T细胞亚群中功能、转录因子、凋亡及干性等相关基因的表达。结果:与效应记忆性T细胞(Tem)和效应性T细胞(Teff)相比,中心记忆性T细胞(Tcm)表面激活性标志物CD69和CD28表达水平增高(P <0. 05),而抑制性分子PD-1表达水平降低(P <0. 05);功能分子(IFN-γ、IL-2)的表达水平上调(P <0. 05);趋化因子受体(CCR7、CXCR4)、STAT3、干性基因(C-myc、Lin28)及抗凋亡等相关基因表达水平上调(P <0. 05)。结论:抗凋亡、自我更新、STAT3等相关基因在Tcm细胞中高表达,且与记忆性T细胞的分化相关,为体外诱导记忆性T细胞提供了基础。Aim:To investigate the differences in phenotype,function and differentiation-related genes expression of different CD8^+T cell subsets.Methods:Peripheral blood mononuclear cells were isolated from healthy human peripheral blood by density gradient centrifugation,and CD8^+T cells were obtained by MACS immunomagnetic beads sorting system.CD8^+T subsets were marked by CD3、CD8、CD45RA and CCR7,and was used to detect the differentiation,phenotype(CD69,CD28,PD-1)and functional molecules(IFN-γ,IL-2)in CD8^+T cells.Flow sorting was used to sort CD8^+T cell subsets.And qRT-PCR was used to detect the expressions of function,transcription factor,apoptosis and stemness-related genes in each differentiation subsets of CD8^+T cells.Results:Compared with effector memory T cells(Tem)and effector T cells(Teff),the expression levels of CD69 and CD28 in central memory T cells(Tcm)were increased(P<0.05),while the expression of PD-1 was decreased(P<0.05).Meanwhile,the expression levels of functional molecules(IFN-γ,IL-2)in Tcm were up-regulated(P<0.05).The results of qRT-PCR showed that the expression levels of chemokine receptors(CCR7,CXCR4),STAT3 gene,the self-renewing genes(C-myc,Lin28),and the anti-apoptotic genes were up-regulated in Tcm(P<0.05).Conclusion:Anti-apoptosis,self-renewal,STAT3 and other related genes are highly expressed in Tcm,and are associated with the differentiation of memory T cells,which provides a basis for inducing memory T cells in vitro.

关 键 词：CD8^+T细胞 分化 记忆性T细胞 中心记忆性T细胞 

分 类 号：R392.12[医药卫生—免疫学]