Arntl基因敲除对小鼠T细胞发育及抗感染功能的影响  

Effect of Arntl on T cell development and anti-infection function in mice

作　　者：孙亚娥 焦安君 王昕[2,3] 张星哲 雷蕾 杨小丰[2,3] 解涛 周晓勃[2,3] 史霖 张保军 刘晓斌[1] SUN Ya’e;JIAO Anjun;WANG Xin;ZHANG Xingzhe;LEI lei;YANG Xiaofeng;XIE Tao;ZHOU Xiaobo;SHI Lin;ZHANG Baojun;LIU Xiaobin(Medical College of Yan’an University,Yan’an 716000;Department of Pathogenic Microbiology and Immunology,School of Basic Medical Sciences,Xi’an Jiaotong University,Xi’an 710061;Institute of Infection and Immunity,Translational Medicine Institute,Xi’an Jiaotong University Health Science Center,Xi’an 710061;Key Laboratory of Environment and Genes Related to Diseases(Xi’an Jiaotong University),Ministry of Education,Xi’an 710061;Xi’an Key Laboratory of Immune Related Diseases,Xi’an 710061,China)

机构地区：[1]延安大学医学院,陕西延安716000 [2]西安交通大学基础医学院病原生物学与免疫学系,陕西西安710061 [3]西安交通大学医学部转化医学研究院感染与免疫研究所,陕西西安710061 [4]环境与疾病相关基因教育部重点实验室(西安交通大学),陕西西安710061 [5]西安市免疫相关疾病重点实验室,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2021年第3期402-407,432,共7页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国际(地区)合作与交流项目(No.81820108017);陕西省免疫相关疾病研究科技创新团队(No.2021TD-38);国家自然科学基金资助项目(No.81771673)。

摘　　要：目的观察Arntl基因敲除对小鼠T细胞发育及抗感染功能的影响。方法构建在T细胞中特异性敲除Arntl基因小鼠(Arntl^(F/F)CD4cre^(+),KO);采用流式细胞术(FCM)检测KO小鼠胸腺和脾脏中T细胞各亚群的比例及数目;检测淋巴细胞脉络丛脑膜炎病毒(lymphocytic choriomeningitis virus,LCMV)感染7 d后小鼠脾脏中T细胞各亚群比例、病毒特异性CD8^(+)T细胞的比例,以及CD8^(+)T细胞中IFN-γ的表达水平;qPCR方法检测小鼠脾脏中病毒载量;借助流式细胞仪分选初始CD4^(+)T细胞(CD4^(+)CD25^(-)CD44^(-)CD62L^(+)T细胞),经体外诱导Th1细胞分化,检测表达IFN-γ细胞的比例。结果与对照组(Arntl^(F/F)CD4cre^(-),WT)小鼠相比,KO小鼠胸腺和脾脏中T细胞各亚群的百分比及数目均无明显改变(P>0.05)。感染LCMV后,KO小鼠脾脏中效应T细胞的比例较对照组无明显改变(P>0.05),但分泌IFN-γT细胞的比例显著增多(P<0.05),病毒清除率升高(P<0.05)。此外,KO小鼠初始CD4^(+)T细胞向Th1细胞分化较对照组显著增强(P<0.05)。结论Arntl在T细胞中特异性敲除不影响T细胞的发育,但会增强T细胞抗病毒感染的能力。Objective To evaluate the effect of Arntl on T cell development and T cell-mediated anti-viral immunity.Methods Arntl^(F/F)CD4cre^(+)(KO)in mice was constructed to delete Arntl gene specifically in T cells.We examined the percentage and number of T cell subsets in the thymus and spleen by flow cytometry(FCM).At day 8 after lymphocytic choriomeningitis virus(LCMV)infection,the proportions of T cell subsets,virus-specific CD8^(+)T cells and IFN-γsecreting T cells were analyzed.The viral load in the spleen was measured using qPCR.Naive CD4^(+)T cells(CD4^(+)CD25^(-)CD44^(-)CD62L^(+))were sorted by flow cytometry to perform T helper cell differentiation in vitro.Results The percentage and number of T cells in the thymus and spleen of KO mice showed no significant change compared with those in the control group(Arntl^(F/F)CD4cre^(-)mice,WT)(P>0.05).Acute LCMV infection did not cause observable changes in effector T cell proportion in the spleen of KO mice compared to that in WT mice(P>0.05),but KO mice showed a higher proportion of IFN-γsecreting T cells(P<0.05)and better virus clearance(P<0.05).In addition,naive CD4^(+)T cells from KO mice were more prone to differentiate into Th1 cells in vitro(P<0.05).Conclusion Arntl deletion in T cells does not affect T cell development,but enhances their ability to defend against viral infection by promoting Th1 cell differentiation and response.

关 键 词：Arntl T细胞发育 病毒感染 免疫应答 辅助性(Th)T细胞分化 

分 类 号：R392[医药卫生—免疫学]