疟原虫感染小鼠肺脏CD4^(+)T细胞中PD-1的表达及功能研究  

Expression of PD⁃1 in CD4^(+) T cells in the lung of Plasmodium infected mice and its functional characteristics

作　　者：蔡琛 陈静 农秀雨 谢安琪 邢俊敏 魏海霞 CAI Chen;CHEN Jing;NONG Xiuyu;XIE Anqi;XING Junmin;WEI Haixia(School of Basic Medical Sciences,Guangzhou Medical University,Guangzhou,Guangdong 511436,China)

机构地区：[1]广州医科大学基础医学院,广东广州511436

出　　处：《热带医学杂志》2024年第2期162-166,264,共6页Journal of Tropical Medicine

基　　金：广东省基础与应用基础研究基金(2021A1515220034);广州市科技计划项目(202102020912);广州医科大学2022年省级大学生创新创业训练计划(2021A093)。

摘　　要：目的探讨宿主感染疟原虫后肺脏CD4^(+)T细胞中程序性细胞死亡1蛋白(PD-1)的表达情况及功能。方法将5~6周雌性C57BL/6小鼠随机分为正常组和感染组,每组6只。采用流式细胞术检测疟原虫感染前后小鼠肺脏CD4^(+)T细胞中PD-1的表达情况,并用流式检测肺脏PD-1+CD4^(+)T细胞表型和细胞因子的分泌能力,之后采用qRT-PCR技术检测可能参与PD-1表达的相关转录因子B淋巴细胞诱导成熟蛋白1(Blimp-1)、活化T细胞核因子(NFATc1)、信号转导及转录激活因子(STAT)3、STAT4、干扰素刺激基因因子3(ISGF3)、激活蛋白1(AP-1)、缺口基因Notch、叉头转录因子1(FoxO1)、核因子-κB(NF-κB)的表达情况。结果疟原虫感染后,小鼠肺脏表达PD-1的CD4^(+)T细胞的比例显著升高,由原来的11.66%升高到58.60%,差异有统计学意义(t=7.892,P<0.01)。感染组小鼠肺脏中PD-1+CD4^(+)T细胞表达CD69分子高于PD-1-CD4^(+)T细胞,差异有统计学意义(t=9.856,P<0.01)。在感染组,与PD-1-CD4^(+)T细胞相比,更多的PD-1+CD4^(+)T细胞分泌IFN-γ、白细胞介素(IL)-2和IL-10,差异均有统计学意义(t=7.301、4.876、7.607,P均<0.05)。qRT-PCR检测结果发现与正常组相比,感染组脾脏中多种转录因子的表达量均改变,差异均有统计学意义(P均<0.05)。结论疟原虫感染后,小鼠肺脏中表达PD-1的CD4^(+)T细胞参与了免疫应答,并通过分泌细胞因子,发挥免疫调节作用。Objective To investigate the expression and function of programmed cell death 1(PD⁃1)on CD4^(+)T cells in lung of mice infected with Plasmodium.Methods Female C57BL/6 mice from 5-6 weeks were randomly divided into normal or infected groups.Flow cytometry was used to detect the expression of PD⁃1 on CD4^(+)T cells with Plasmodium infection,and the phenotype and cytokine secreting ability of CD4^(+)T cells expressing PD⁃1 was detected by flow cytometry.qRT⁃PCR was used to detect the expression of related transcription factors B lymphocyte⁃induced maturation protein 1(Blimp⁃1),nuclear factor⁃activated T cell 1(NFATc1),signal transducer and activator of transcription(STAT)3,STAT4,interferon⁃stimulated gene factor 3(ISGF3),activator protein 1(AP⁃1),Notch,forkhead box transcription factor 1(FoxO1),nuclear factor⁃κB(NF⁃κB).Results After infection with Plasmodium,the proportion of CD4^(+)T cells expressing PD⁃1 in the lungs of mice was increased significantly from 11.66%to 58.60%,the difference was statistically significant(t=7.892,P<0.01).In the infected group,PD⁃1+CD4^(+)T cells expressed more CD69 than PD⁃1-CD4^(+)T cells,which showed a statistically significant difference(t=9.856,P<0.01).In the infected group,compared with PD⁃1-CD4^(+)T cells,more PD⁃1+CD4^(+)T cells secreted IFN⁃γ,interleukin(IL)⁃2 and IL⁃10,the differences were statistically significant(t=7.301,4.876,7.607;all P<0.05).QRT⁃PCR showed that compared with the normal group,the expression level of various transcription factors was changed in the infected group,which showed a statistically significant difference(all P<0.05).Conclusion After infected with Plasmodium,CD4^(+)T cells expressed PD⁃1 in the lung of mice were involved in the immune response,and played a significant role in immunomodulatory through secreting cytokines.

关 键 词：疟原虫 肺脏 CD4^(+)T细胞 程序性细胞死亡1蛋白 

分 类 号：R382.31[医药卫生—医学寄生虫学]