多房棘球蚴感染小鼠腹膜腔免疫细胞迁移及其功能变化  

Immune cell migration and its functional changes in the peritoneal cavity of mice infected with Echinococcus multilocular

作　　者：邓冰清 阿迪莱·多力坤 李寅时 阿比旦·艾尼瓦尔 孙胜 肖雯颖 葛聪蕙 唐娜 姜涛[3] 王慧 张传山[1,2] 李静 DENG Bingqing;ADILAI Duolikun;LI Yinshi;ABIDAN Ainiwaer;SUN Sheng;XIAO Wenying;GE Conghui;TANG Na;JIANG Tao;WANG Hui;ZHANG Chuanshan;LI Jing(College of Basic Medical,Xinjiang Medical University,Urumqi 830054,Xinjiang,China;Clinical Medicine Research Institute,the First Afilicated Hospital of Xinjiang Medical University,Urumqi 830054,Xinjiang,China;Xinjiang Medical University Laboratory Animal Center,Urumqi 830011,Xinjiang,China;Key Laboratory of High Incidence Disease Research in Xingjiang(Xinjiang Medical University),Ministry of Education,Urumqi 830054,Xinjiang,China;National Institute of Parasitic Diseases,Chinese Center for Disease Control and Prevention(Chinese Center for Tropical Diseases Research),NHC Key Laboratory of Parasite and Vector Biology(National Institute of Parasitic Diseases,Chinese Center for Disease Control and Prevention),Shanghai 200025,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,新疆乌鲁木齐830054 [3]新疆医科大学实验动物中心,新疆乌鲁木齐830011 [4]新疆地区高发疾病研究教育部重点实验室(新疆医科大学),新疆乌鲁木齐830054 [5]中国疾病预防控制中心寄生虫病预防控制所(国家热带病研究中心),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(中国疾病预防控制中心寄生虫病预防控制所),上海200025

出　　处：《中国寄生虫学与寄生虫病杂志》2025年第1期76-83,共8页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：新疆维吾尔自治区自然科学基金(2022D01E51,2022D01D60);国家自然科学基金(82302561,82372279,82160396);新疆地区高发疾病研究教育部重点实验室开放课题资助项目(2023B03);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-2024-25);国家卫健委寄生虫病原与媒介生物学重点实验室开放课题资助项目(NHCKFKT2022-05)。

摘　　要：目的了解多房棘球蚴感染小鼠腹膜腔免疫细胞迁移及分化的变化,探讨其发挥免疫功能的机制。方法裂解增强绿色荧光蛋白(EGFP)小鼠脾组织,分离EGFP示踪免疫细胞。C57BL/6J小鼠腹腔注射2.5×10^(7)个示踪免疫细胞,5 d后经肝门静脉穿刺接种3000个多房棘球蚴原头节。感染19周后,取感染小鼠肝、脾组织,冷冻切片后4,6-二脒基-2-苯基吲哚(DAPI)染色,免疫荧光观察示踪免疫细胞的分布。分离感染小鼠外周血、肝、脾和腹膜腔免疫细胞,流式细胞术检测EGFP示踪免疫细胞在不同组织的迁移分布和细胞分型,以及细胞因子γ干扰素(IFN-γ)、肿瘤坏死因子-α(TNF-α)、白细胞介素4(IL-4)的分泌情况。结果免疫荧光结果显示,EGFP示踪细胞主要迁移至小鼠脾组织。肝组织中各型示踪淋巴细胞占比从高到低依次为CD8^(+)T细胞(79.63±5.00)%、CD4^(+)T细胞(7.08±4.00)%、B细胞(2.01±1.00)%(F=396.20,P<0.01);脾组织中各型淋巴细胞占比从高到低依次为CD8^(+)T细胞(73.97±3.56)%、CD4^(+)T细胞(11.92±5.02)%、B细胞(3.86±0.32)%(F=349.00,P<0.01);腹膜腔中各型淋巴细胞占比从高到低依次为CD8^(+)T细胞(45.27±4.53)%、CD4^(+)T细胞(34.60±8.00)%、B细胞(15.03±6.38)%(F=16.90,P<0.05)。肝、脾、腹膜腔中的示踪CD4^(+)T细胞数量分别为2700±3120、14407±11958、3376±1481,示踪CD8^(+)T细胞数量分别为26407±22190、109179±92692、4245±798,示踪B细胞数量分别为698±719、5794±4971、1476±840,3种示踪淋巴细胞均主要分布在脾组织中(F=2.51、3.03、2.62,均P>0.05)。肝组织中各型髓系细胞占比从高到低依次为树突状细胞(3.94±1.33)%、单核细胞(3.79±1.80)%、巨噬细胞(3.13±1.85)%、嗜酸粒细胞(2.40±1.81)%、骨髓来源抑制性细胞(0.76±0.17)%、中性粒细胞(0.67±0.04)%(F=3.21,P<0.05);脾组织中各型髓系细胞占比从高到低依次为树突状细胞(1.86±0.89)%、单核细胞(0.41±0.09)%、巨噬细胞(0.23±0.03)%、中�Objective To understand the changes in the migration and differentiation of immune cells in the peritoneal cavity of mice infected with Echinococcus multilocularis,and to explore the mechanism of immune function.Methods Spleen tissues of enhanced green fluorescent protein(EGFP)mice were lysed to isolate EGFP-labeled im-mune cells.C57BL/6J mice were intraperitoneally injected with 2.5×10^(7)traced immune cells.Five days later,3000 protoscoleces of E.multilocularis were inoculated via the hepatic portal vein.After 19 weeks of infection,the liver and spleen tissues of the infected mice were collected.After frozen sectioning,the sections were stained with 4,6-di-amidino-2-phenylindole(DAPI),and immunofluorescence was used to observe the distribution of the traced immune cells.Immune cells from peripheral blood,liver,spleen and peritoneal cavity of infected mice were isolated,and the migration distribution and cell typing of EGFP-labeled immune cells in different tissues were detected by flow cytometry,as well as the secretion of cytokines interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α)and interleukin-4(IL-4).Results The immunofluorescence results showed that the EGFP-traced cells mainly migrated to the spleen tissue of the mice.In the liver tissue,the proportions of various types of traced lymphocytes from high to low were as follows:CD8^(+)T cells(79.63±5.00)%,CD4^(+)T cells(7.08±4.00)%,and B cells(2.01±1.00)%(F=396.20,P<0.01).In the spleen tissue,the proportions of various types of lymphocytes from high to low were as follows:CD8^(+)T cells(73.97±3.56)%,CD4^(+)T cells(11.92±5.02)%and B cells(3.86±0.32)%(F=349.00,P<0.01).In the peritoneal cavity,the propor-tions of various types of lymphocytes from high to low were as follows:CD8^(+)T cells(45.27±4.53)%,CD4^(+)T cells(34.60±8.00)%and B cells(15.03±6.38)%(F=16.90,P<0.05).The numbers of traced CD4^(+)T cells in the liver,spleen,and peritoneal cavity were 2700±3120,14407±11958 and 3376±1481 respectively.The numbers of traced CD8^(+)T cells were 2640

关 键 词：多房棘球蚴病 腹膜腔 免疫细胞 迁移 CD8^(+)T细胞 

分 类 号：R532.32[医药卫生—内科学]