旋毛虫胶质瘤致病相关蛋白1样蛋白2对宿主免疫功能的影响  

Effects of glioma pathogenesis-related protein 1-like protein 2 fromTrichinella spiralis on host immune functions

作　　者：孙晓轲 陆滢滢 齐萌[2] 陈伟[2] 陆明敏 徐立新[1] 宋小凯[1] 李祥瑞[1] 严若峰[1] SUN Xiaoke;LU Yingying;QI Meng;CHEN Wei;LU Mingmin;XU Lixin;SONG Xiaokai;LI Xiangrui;YAN Ruofeng(College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China;College of Animal Science,Tarim University,Alar 843300,China)

机构地区：[1]南京农业大学动物医学院,江苏南京210095 [2]塔里木大学动物科学学院,新疆阿拉尔843300

出　　处：《南京农业大学学报》2023年第4期727-737,共11页Journal of Nanjing Agricultural University

基　　金：江苏省自然科学基金项目(BK20141365);新疆生产建设兵团重点领域科技攻关计划项目(2020AB025);南京农业大学SRT计划项目(202017XX19)。

摘　　要：[目的]本文旨在分析旋毛虫胶质瘤致病相关蛋白1样蛋白2(GLIPR1L2)对宿主免疫功能的影响。[方法]用反转录PCR(RT-PCR)方法克隆旋毛虫GLIPR1L2基因,构建表达载体,获得重组蛋白(rGLIPR1L2),用Western blot分析rGLIPR1L2的反应原性。将大鼠外周血单核细胞(PBMC)与rGLIPR1L2共孵育,用免疫荧光检测技术(IFA)检测rGLIPR1L2与大鼠PBMC的结合,分析rGLIPR1L2对细胞增殖、迁移、NO分泌、吞噬功能和细胞凋亡的影响。大鼠腹腔注射rGLIPR1L2后,测定血清中IL-4、IL-9、IL-17、TGF-β、IFN-γ等细胞因子的变化。背部皮下多点注射对小鼠进行免疫后,ELISA检测特异性IgG、IgG1和IgG2a抗体的变化,收集旋毛虫成虫和肌肉幼虫并计数,分析rGLIPR1L2的免疫保护作用。[结果]体外试验表明,rGLIPR1L2能够促进大鼠PBMC的增殖与迁移,增强细胞NO分泌和吞噬功能,一定浓度下可提高细胞凋亡比例。体内试验显示rGLIPR1L2能促进大鼠IL-4分泌,但对细胞因子IFN-γ、IL-9、IL-17和TGF-β的分泌无显著影响。rGLIPR1L2免疫小鼠可引起其体内特异性IgG抗体水平显著升高,且可显著减少受感染小鼠的肌肉幼虫数。[结论]旋毛虫胶质瘤致病相关蛋白1样蛋白2能通过多种途径影响宿主免疫功能。[Objectives]This research aimed to analyze the effect of Trichinella spiralis glioma pathogenesis-related protein 1-like protein 2(GLIPR1L2)on host immune functions.[Methods]The gene of Trichinella spiralis GLIPR1L2 was cloned by reverse transcript PCR.The open reading frame of this gene was subcloned into prokaryotic expression vector,and the recombinant protein(rGLIPR1L2)was obtained.The reactogenicity of rGLIPR1L2 was analyzed by Western blot.Rat peripheral blood mononuclear cells(PBMC)were incubated with rGLIPR1L2.The binding of rGLIPR1L2 to rat PBMC was observed.The effects on cell proliferation,migration,NO production,phagocytosis and apoptosis were analyzed.The changes of IL-4,IL-9,IL-17,TGF-β,IFN-γand other cytokines in serum were measured after rats were intraperitoneal injected with rGLIPR1L2.To study the protective efficiency,mice were immunized with rGLIPR1L2.The changes of specific IgG,IgG1 and IgG2a antibodies were detected by ELISA.The adult worms and muscle larvae were collected and counted.[Results]In vitro experiments showed that rGLIPR1L2 could promote the proliferation and migration of rat PBMC,enhance the NO secretion and phagocytosis,and increase the proportion of apoptosis at a certain concentration.In vivo experiments showed that rGLIPR1L2 could promote the secretion of IL-4 in rats,but had no significant effect on the secretion of IFN-γ,IL-9,IL-17 and TGF-β.Mice vaccinated with rGLIPR1L2 resulted in a significant increase in specific IgG antibodies and a significant reduction in the number of muscle larva.[Conclusions]Trichinella spiralis glioma pathogenesis-related protein 1-like protein 2 can affect the host immune function through various ways.

关 键 词：旋毛虫 胶质瘤致病相关蛋白1样蛋白2 外周血单核细胞 免疫保护 

分 类 号：S852.73[农业科学—基础兽医学]