多房棘球蚴感染对小鼠脾脏自然杀伤细胞Tim 3表达的影响  

Effect of Echinococcus multilocularis infection on Tim3 expression in spleen natural killer cells of mice

作　　者：施阳 阿比旦·艾尼瓦尔 李德伟 孜比姑·肉素 王茂林 郑旭然 康雪娇 王慧 李静[1,2] 张传山 SHI Yang;ABIDAN Ainiwaer;LI Dewei;ZIBIGU Rousu;WANG Maoling;ZHENG Xuran;KANG Xuejiao;WANG Hui;LI Jing;ZHANG Chuanshan(College of Basic Medical Sciences,Xinjiang Medical University,Urumqi,Xinjiang 830054,China;Institute of Clinical Medi-cine,The First Affiliated Hospital of Xinjiang Medical University,State Key Laboratory of Pathogenesis,Prevention and Treatment f High Incidence Diseases in Central Asia,Xinjiang Uygur Autonomous Region Key Laboratory of Echinococcosis,Urumqi,Xinji-ang 830054,China;Xinjiang Uygur Autonomous Region Key Laboratory of Molecular Biology for Endemic Diseases,China)

机构地区：[1]新疆医科大学基础医学院,新疆乌鲁木齐830054 [2]新疆医科大学第一附属医院临床医学研究院、省部共建中亚高发病成因与防治国家重点实验室、新疆维吾尔自治区包虫病基础医学重点实验室,新疆乌鲁木齐830054 [3]新疆维吾尔自治区地方病分子生物学重点实验室

出　　处：《中国血吸虫病防治杂志》2023年第4期366-373,共8页Chinese Journal of Schistosomiasis Control

基　　金：国家重点研发计划项目(2021YFC2300800,2021YFC2300801);国家自然科学基金(82160396,81760368,82060370);国家杰出青年科学基金培育项目(xyd2021J003);新疆维吾尔自治区自然科学基金(2022D01D60,2022D01E51);新疆维吾尔自治区青年科技拔尖人才项目(2022TSYCCX0106);省部共建中亚高发病成因与防治国家重点实验室开放课题资助项目(SKL-HIDCA-2021-11);国家卫生健康委员会寄生虫病原与媒介生物学重点实验室开放课题(NHCKFKT2022-05)。

摘　　要：目的探讨多房棘球蚴感染对小鼠脾脏自然杀伤(natural killer,NK)细胞Tim3及其与免疫检查点分子2B4和LAG3共表达的影响。方法取体质量为(20±2)g的C57BL/6小鼠,随机分为高剂量感染组(15只)、低剂量感染组(13只)和对照组(11只),高、低剂量感染组分别经肝门静脉接种2000、50个多房棘球蚴原头节,对照组经肝门静脉注射等量生理盐水。分别于感染后12、24周收集小鼠脾脏细胞,采用流式细胞术检测小鼠脾脏NK细胞Tim3表达及其与2B4和LAG3共表达水平。结果多房棘球蚴感染后12周,各组小鼠脾脏NK细胞Tim3表达比例及其与2B4共表达比例差异有统计学意义(F=13.559、12.465,P均<0.01);低剂量感染组小鼠脾脏NK细胞Tim3表达比例[(23.84±2.28)%]高于高剂量感染组[(15.72±3.67)%]和对照组[(16.14±3.83)%](P均<0.01);低剂量感染组小鼠脾脏NK细胞Tim3和2B4共表达比例[(22.20±2.13)%]高于高剂量感染组[(14.17±3.81)%]和对照组[(15.20±3.77)%](P均<0.01)。多房棘球蚴感染后24周,各组小鼠脾脏NK细胞Tim3表达比例及其与2B4共表达比例差异有统计学意义(F=5.243、4.659,P均<0.05);高剂量感染组小鼠脾脏NK细胞Tim3表达比例[(20.55±7.04)%]、Tim3和2B4共表达比例[(20.98±7.12)%]均低于对照组[(31.38±3.19)%、(31.25±3.06)%](P均<0.05),低剂量感染组小鼠脾脏NK细胞Tim3表达比例[(26.80±6.47)%]、Tim3和2B4共表达比例[(26.48±6.48)%]与对照组差异均无统计学意义(P均>0.05)。感染后12、24周,各组小鼠脾脏NK细胞Tim3和LAG3共表达比例差异均无统计学意义(F=2.283、0.375,P均>0.05)。低剂量感染组中,多房棘球蚴感染后12、24周小鼠脾脏NK细胞Tim3表达比例、Tim3和2B4共表达比例差异均无统计学意义(t=-1.137、-1.658,P>0.05);感染后24周,小鼠脾脏NK细胞Tim3和LAG3共表达比例较感染后12周上调(t=-5.261,P<0.01)。高剂量感染组中,多房棘球蚴感染后12、24周小鼠脾脏NK细胞Tim3表达比例差异无统计�Objective To investigate the effect of Echinococcus multilocularis infection on Tim3 expression and its co-expression with immune checkpoint molecules 2B4 and LAG3 in spleen natural killer(NK) cells of mice.Methods C57BL/6 mice,each weighing(20±2) g,were randomly divided into a high-dose infection group(15 mice),a low-dose infection group(13 mice),and a control group(11 mice).Mice in the high-and low-dose infection groups were inoculated with 2 000 and 50 Echinococcus multilocularis protoscolices via the hepatic portal vein,while animals in the control group was injected with an equivalent amount of physiological saline via the hepatic portal vein.Mouse spleen cells were harvested 12 and 24 weeks post-infection,and Tim3expression and its co-expression with 2B4 and LAG3 in NK cells were detected using flow cytometry.Results There were significant differences in the proportions of Tim3 expression(F=13.559,P<0.001) and Tim3 and 2B4 co-expression(F=12.465,P<0.001) in mouse spleen NK cells among groups 12 weeks post-infection with E.multilocularis,and the proportion of Tim3 expression was significantly higher in mouse spleen NK cells in the low-dose infection group [(23.84±2.28)%] than in the highdose infection group [(15.72±3.67)%] and the control group [(16.14±3.83)%](both P values 0.01),while the proportion of Tim3 and 2B4 co-expression was significantly higher in mouse spleen NK cells in the low-dose infection group [(22.20±2.13)%]than in the high-dose infection group [(14.17±3.81)%] and the control group [(15.20±3.77)%](both P values 0.01).There were significant differences in the proportions of Tim3 expression(F=5.243,P<0.05) and Tim3 and 2B4 co-expression(F=4.659,P<0.05) in mouse spleen NK cells among groups 24 weeks post-infection with E.multilocularis infection,and the proportions of Tim3 expression and Tim3 and 2B4 co-expression were significantly lower in mouse spleen NK cells in the high-dose infection group [(20.55±7.04)% and(20.98±7.12)%] than in the control group [(31.38±3.19)% and(31.25±3.06

关 键 词：多房棘球蚴 脾脏细胞 Tim3 自然杀伤细胞 免疫检查点 

分 类 号：R383.33[医药卫生—医学寄生虫学]