表面展示柔嫩艾美耳球虫EtMIC2蛋白的侵入型乳酸菌的免疫特性研究  

Immune properties of invasive lactic acid bacteria expressing EtMIC2 protein of Eimeria tenella on its surface

作　　者：张赞 刘晶 高兴 刘洋 姜延龙 王春凤 杨桂连 ZHANG Zan;LIUJing;GAO Xing;LIU Yang;JIANG Yan-long;WANG Chun-feng;YANG Gui-lian(Key Laboratory of Animal Production and Quality Security,Ministry of Education/Jilin Provincial Engineering Research Center of Animal Probiotics/College of Animal Science and Technology ,Jilin Agricultural University, Changchun 130118,China)

机构地区：[1]吉林农业大学动物科学技术学院吉林省动物微生态制剂工程研究中心动物生产及产品质量安全教育部重点实验室,吉林长春130118

出　　处：《中国兽医科学》2019年第5期619-624,共6页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2017YFD 0501200);国家自然科学基金项目(31672528);吉林省科技发展计划项目(20160519011JH;20170204034N Y;20180201040NY)

摘　　要：为了研究表达pgsA′-EtMIC2融合蛋白的侵入型乳酸菌的免疫特性,将pgsA′-EtMIC2基因片段插入到载体pSIP409-FnBPA中,得到pSIP409-FnBPA-pgsA′-EtMIC2重组质粒,并用电转化的方式转入植物乳杆菌NC8感受态细胞中,构建表面展示EtMIC2蛋白的侵入型乳酸菌。通过检测免疫雏鸡外周血淋巴细胞中CD3^+CD4^+、CD3^+CD8^+T淋巴细胞的发育情况,血清中EtMIC2抗体、IL-4、IL-18、IFN-γ的含量以及肠道灌洗液中IgA的分泌量来评价该侵入型乳酸菌对雏鸡的免疫特性。结果显示,成功构建出携带pSIP409-FnBPA-pgsA′-EtMIC2质粒的重组乳酸菌;雏鸡免疫后外周血淋巴细胞中CD3^+CD4^+、CD3^+CD8^+T淋巴细胞含量明显升高;EtMIC2抗体、IL-18、IFN-γ细胞因子及IgA产生的水平均显著上升,表明该侵入型乳酸菌具有一定的免疫效力,为后期研究抗球虫的免疫保护效果奠定了基础。In order to study the immune properties of the invasive lactic acid bacteria expressing the pgs A’-Et MIC2,the gene fragment was inserted into the p SIP409-Fn BPA to obtain the pSIP409-Fn BPApgs A’-Et MIC2,and was transfered into Lactobacillus plantarum NC8 by electroporation to construct an invasive lactic acid bacteria displaying Et MIC2 protein of Eimeria tenella on its surface. After immunization with the recombinant bacteria,its influence on T lymphocytes was detected by flow cytometry,and the expression of Et MIC2,IL-4,IL-18,IFN-γ and s Ig A was also tested by ELISA. The result showed that recombinant lactic acid bacteria carrying the p SIP409-Fn BPA-pgs A’-Et MIC2 plasmid were constructed successfully. Compared with the control group,the contents of CD3^+CD4^+ and CD3^+CD8^+T lymphocytes were significantly higher and the expression of anti-Et MIC2,IL-18,IFN-γ in serum were increased,alsos IgA in intestinal lavage fluid. Together,the recombinant bacteria,to some degree,could provide immune effects,and laid the foundation for the evaluation of the immune protection against coccidiosis.

关 键 词：柔嫩艾美耳球虫 EtMIC2 FnBPA 侵入型乳酸菌 免疫特性 

分 类 号：S852.723[农业科学—基础兽医学]