多房棘球蚴感染小鼠脾CD4^+T细胞亚群及其功能耗竭的变化  

Changes in subsets and functional exhaustion of CD4^+T cells in spleens of mice infected with Echinococcus multilocularis

作　　者：侯昕伶 李玲慧 李亮 李静[2] 王慧 邵英梅 张传山 HOU Xin-ling;LI Ling-hui;LI Liang;LI Jing;WANG Hui;SHAO Ying-mei;ZHANG Chuan-shan(Clinical Medicine Institute,State Key Laboratory of Pathogenesis,Prevention and Treatment of High Incidence Diseases in Central Asia,Xinjiang Key Laboratory of Echinococcosis,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 852204,China;Basic Medical College,Xinjiang Medical University,Urumqi 852204,China;College of Animal Medicine,Xinjiang Agricultural University,Urumqi 852204,China;Department of Hepatic Hydatid and Hepatobiliary Surgery,Digestive and Vascular Surgery Centre,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 852204,China)

机构地区：[1]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,新疆包虫病基础医学重点实验室,乌鲁木齐830054 [2]新疆医科大学基础医学院,乌鲁木齐830054 [3]新疆农业大学动物医学学院,乌鲁木齐830054 [4]新疆医科大学第一附属医院,消化血管外科中心肝胆包虫科,乌鲁木齐830054

出　　处：《中国寄生虫学与寄生虫病杂志》2020年第5期611-618,624,共9页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：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(No.SKL-HIDCA-2019-2);国家自然科学基金(No.81760368,No.81560330);新疆维吾尔自治区高校科研计划项目(No.XJEDU2020Y024)。

摘　　要：目的研究不同数量多房棘球蚴感染对小鼠脾CD4^+T细胞亚群及其免疫功能的影响。方法60只C57BL/6小鼠随机分为4组,每组15只,分别为假手术组、低数量感染组(50个原头节)、中数量感染组(500个原头节)和高数量感染组(2 000个原头节)。小鼠麻醉后经肝门静脉部位穿刺,注射不同数量原头节,假手术组注射等量生理盐水。于感染后2、 12和24周各组分别取5只小鼠,取脾组织研磨分离淋巴细胞。流式细胞术检测各组小鼠脾CD4^+T细胞记忆表型、不同亚群比例、免疫抑制性分子淋巴细胞活化蛋白3 (LAG3)表达。采用GraphPad Prism 6.0软件进行作图和统计学分析。结果感染后2周,低数量和中数量感染组小鼠脾CD4^+IFN-γ^+T细胞比例分别为(7.54±1.44)%、(7.58±3.17)%,高于假手术组的(3.52±1.03)%(P <0.05);CD4^+TNF-α^+T细胞比例分别为(39.34±4.19)%、(39.53±10.74)%,高于假手术组(22.62±1.50)%(P <0.01)。感染后12周,低数量和中数量感染组小鼠脾CD4^+IFN-γ^+T细胞比例分别为(16.52±0.77)%、(22.98±4.32)%,高于假手术组(16.88±2.49)%(P <0.05);CD4^+TNF-α^+T细胞比例分别为(27.26±2.12)%、(28.36±5.24)%,高于假手术组(19.72±3.87)%(P <0.05);CD4^+IL17A^+T细胞比例分别为(10.70±1.81)%、(11.52±2.68)%,高于假手术组(5.40±1.32)%(P <0.01);同时,低数量和中数量感染组小鼠脾CD4^+IL-4^+T细胞比例分别为(2.87±0.84)%、(3.50±0.77)%,高于假手术组(1.75±0.83)%(P <0.01);CD4^+IL-10^+T细胞比例分别为(4.63±0.78)、(7.09±2.42)%,高于假手术组(3.03±0.79)%(P <0.01)。感染后24周,中数量、高数量感染组小鼠脾CD4^+IFN-γ^+T、 CD4^+TNF-α^+T、 CD4^+IL-4^+T、 CD4^+IL-10^+T和CD4^+IL17A^+T细胞的比例均高于假手术组(P <0.05),且高数量组小鼠脾Treg细胞的比例高于假手术组(P <0.01),各感染组小鼠脾效应记忆性CD4^+T细胞比例高于假手术组;各感染组小鼠脾CD4^+LAG3^+T细胞比例分别为(16.45±4.89)%、(14.54±4.96)%、(14.62±2.43Objective To investigate the effects of different doses of Echinococcus multilocularis infection on splenic CD4^+T cell subsets and immune functions in mice.Methods Sixty female C57 BL/6 mice were randomly divided into 4 groups,15 in each group,including sham operation group(saline),low-dose infection group(50 protoscoleces/mouse),medium-dose infection group(500 protoscoleces/mouse),and high-dose infection group(2000 protoscoleces/mouse).Mice were inoculated via the hepatic portal vein under anaesthetia with different doses of protoscoleces in saline,whereas the control mice were injected with the same volume of saline.Spleens were taken from 5 mice of each group at 2,12,and 24 weeks after infection,and ground to isolate lymphocytes.Flow cytometry was performed to detect the memory phenotype,the proportions of different subsets,and LAG3 expression in splenic CD4^+T cells in different experimental groups.Statistical analysis was performed using Graphad Prism 6.0 software.Results At 2 weeks after infection,the proportions of splenic CD4^+IFN-γ^+T cells in the low-and mediumdose groups were(7.54±1.44)%and(7.58±3.17)%,respectively,which were higher than that in the sham operation group[(3.52±1.03)%,P<0.05];and the proportions of splenic CD4^+TNF-α^+T cells were(39.34±4.19)%and(39.53±10.7)%,respectively,which were higher than that of the sham operation group[(22.62±1.50)%,P<0.01].At12 weeks after infection,the proportions of splenic CD4^+IFN-γ^+T cells in the low-and medium-dose groups were(16.52±0.77)%and(22.98±4.32)%,respectively,which were higher than that in the sham operation group[(16.88±2.49)%,P<0.05];the proportions of splenic CD4^+TNF-α^+T cells were(27.26±2.12)%and(28.36±5.24)%,respectively,which were higher than that of the sham operation group[(19.72±3.87)%,P<0.05];and the proportions of splenic CD4^+IL-17 A^+T cells were(10.70±1.81)%and(11.52±2.68)%,respectively,which were higher than that of the sham operation group[(5.40±1.32)%,P<0.01].In addition,the proportions of splenic CD4^+IL-4^

关 键 词：多房棘球蚴病 脾 CD4^+T细胞 功能耗竭 淋巴细胞活化蛋白3 

分 类 号：R532.32[医药卫生—内科学]