多房棘球蚴感染小鼠外周血中髓源抑制性细胞比例动态变化及细胞因子表达研究  

Study on the dynamic change of myeloid⁃derived suppressor cells proportion and cytokine expression in peripheral blood of mice infected with Echinococcus multilocularis

作　　者：苏雅馨 江楠 章孝成 王莹 蒋小凤 霍乐乐 王雅雪 曹建平 沈玉娟 SU Yaxin;JIANG Nan;ZHANG Xiaocheng;WANG Ying;JIANG Xiaofeng;HUO Lele;WANG Yaxue;CAO Jianping;SHEN Yujuan(National Institute of Parasitic Diseases,Chinese Center for Disease Control and Prevention,Chinese Center for Tropical Diseases Research,National Key Laboratory of Intelligent Tracking andForecasting for Infectious Diseases,Key Laboratory on Parasite and Vector Biology,Ministry of Health,WHO Collaborating Centre for Tropical Diseases,National Center for International Research on Tropical Diseases,Ministry of Science and Technology,Shanghai 200025,China;School of Global Health,Chinese Center for Tropical Diseases Research,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China)

机构地区：[1]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),传染病溯源预警与智能决策全国重点实验室,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,世界卫生组织热带病合作中心,科技部国家级热带病国际研究中心,上海200025 [2]上海交通大学医学院-国家热带病研究中心全球健康学院,上海200025

出　　处：《中国寄生虫学与寄生虫病杂志》2024年第2期211-216,共6页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金(82372283,82072307)。

摘　　要：目的分析多房棘球蚴原头节感染小鼠外周血白细胞中髓源抑制性细胞(MDSC)及其亚型的比例动态变化,以及感染晚期小鼠血清中与MDSC增殖相关细胞因子的表达情况。方法将24只BALB/c小鼠随机分为感染组和对照组,每组12只。感染组小鼠经腹腔注射1200个原头节,对照组小鼠注射等量生理盐水。每组小鼠分别于感染后30、90和180 d随机各取3只,观察肝、脾组织形态学变化,并采集眼眶静脉丛外周血制备白细胞。外周血白细胞用外标抗体CD11b、Gr-1、Ly6G和Ly6C孵育后,流式细胞仪检测MDSC及其亚型多核MDSC(PMN-MDSC)和单核MDSC(M-MDSC)的占比。感染后180 d,采集感染组和对照组小鼠血清,ELISA检测白细胞介素6(IL-6)、IL-13、肿瘤坏死因子α(TNF-α)、粒细胞-巨噬细胞集落刺激因子(GM-CSF)的浓度。使用GraphPad Prism 9.0软件进行制图与统计学分析,两组间比较采用独立样本t检验。结果感染多房棘球蚴原头节后30 d,感染组小鼠肝组织出现囊泡,囊泡的体积随感染时间延长而增加;脾组织随感染时间延长逐渐肿大。流式细胞分析结果显示,感染后30、90和180 d,感染组小鼠外周血白细胞中MDSC占比分别为(13.2±2.4)%、(15.7±2.3)%和(41.3±4.0)%,对照组小鼠的占比分别为(12.4±3.2)%、(6.0±0.9)%和(22.3±1.1)%,感染后90和180 d感染组均高于对照组(t=3.949、4.682,P<0.05、0.01);感染后30、90和180 d,感染组小鼠外周血白细胞中PMN-MDSC占比分别为(10.9±2.1)%、(12.5±2.4)%和(35.8±3.6)%,对照组小鼠的占比分别为(9.6±3.1)%、(4.5±0.6)%和(18.5±0.6)%,感染后90和180 d感染组均高于对照组(t=3.237、4.788,P<0.05、0.01);感染后30、90和180 d,感染组小鼠外周血白细胞中M-MDSC占比分别为(1.8±0.3)%、(1.1±0.1)%和(4.6±1.1)%,对照组小鼠的占比分别为(2.3±0.2)%、(0.5±0.1)%和(3.4±0.9)%,感染后90 d感染组高于对照组(t=3.246,P<0.05)。感染后180 d,感染组小鼠血清中IL-6、IL-13、TNF-α�Objective To analyze the dynamic changes in the proportion of myeloid-derived suppressor cells(MDSCs)and their subtypes in peripheral blood leukocytes of mice infected with Echinococcus multilocularis protoscoleces and detect the expression of cytokines related to MDSCs proliferation in the late-infected mice serum.Meth⁃ods Twenty-four BALB/c mice were randomly divided into the infected group and the control group,with 12 mice in each group.Mice in the infected group were intraperitoneally injected with 1200 E.multilocularis protoscoleces,while mice in the control group injected with the same volume of normal saline.Three mice from each group were randomly selected at 30,90 and 180 days after infection to observe histomorphological changes in liver and spleen.Peripheral blood was collected from orbital venous plexus to prepare peripheral blood leukocytes.After incubation with externally labeled antibodies to CD11b,Gr-1,Ly6G and Ly6C,flow cytometry was used to detect the proportions of MDSCs,polymorphonuclear MDSCs(PMN-MDSCs)and monocytic MDSCs(M-MDSCs).Serum samples from both groups were collected 180 days after infection for determine the concentrations of interleukin-6(IL-6),IL-13,tumor necrosis factorα(TNF-α)and granulocyte-macrophage colony-stimulating factor(GM-CSF)by ELISA assay.GraphPad Prism 9.0 software was utilized for graphical and statistical analysis;independent sample t-test was used for intergroup comparison.Results After E.multilocularis protoscoleces infection,cysts appeared in the liver of infected group mice 30 days after infection,and volume of cysts was increased over time.Spleen of infected group mice were enlaged over time 30,90,and 180 days after infection.The results of flow cytometry showed that 30,90 and 180 days after infection,the proportion of MDSCs in peripheral blood leukocytes of infected group were(13.2±2.4)%,(15.7±2.3)%and(41.3±4.0)%,respectively.And the proportion of MDSCs in the control group were(12.4±3.2)%,(6.0±0.9)%and(22.3±1.1)%,respectively.The infected group wa

关 键 词：多房棘球蚴 原头节 髓源抑制性细胞 细胞因子 

分 类 号：R532.32[医药卫生—内科学]