猪带绦虫丝氨酸蛋白酶抑制剂Ts-serpin-1对人巨噬细胞THP-1的免疫调节功能研究  

Study on immunomodulatory function of Taenia solium serine protease inhibitorTs-serpin-1 in THP-1 cells

作　　者：毕研丽[1] 刘仲藜[1,2] 郭爱疆[1] 张少华[1] 王帅[1] 才学鹏[1,3] BI Yan-li;LIU Zhong-li;GUO Ai-jiang;ZHANG Shao-hua;WANG Shuai;CAI Xue-peng(State Key Laboratory of Veterinary Etiological Biology/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China;College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China;China Institute of Veterinary Drug Control,Beijing 100081,China)

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室,甘肃兰州730046 [2]甘肃农业大学动物医学院,甘肃兰州730070 [3]中国兽医药品监察所,北京100081

出　　处：《中国兽医科学》2021年第2期161-168,共8页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31802179,31772726);国家重点研发计划项目(2017YFD0501303)。

摘　　要：主要研究猪带绦虫丝氨酸蛋白酶抑制剂Ts-serpin-1(WormBase:TsM_000065700)对宿主THP-1细胞的免疫调节作用。通过设计特异性引物和RT-PCR扩增技术,获得Ts-serpin-1编码序列,用qRT-PCR分析Ts-serpin-1基因在猪带绦虫成虫和中绦期幼虫的表达情况;构建pCold-Ts-serpin-1原核表达载体,诱导表达纯化重组蛋白Ts-serpin-1;用重组蛋白Ts-serpin-1处理THP-1细胞,采用qRT-PCR和ELISA方法检测Ts-serpin-1处理THP-1细胞后,各炎性细胞因子的变化情况。结果显示:获得的Ts-serpin-1目的基因长度为1149 bp,编码382个氨基酸,含有Serpin家族特有的反应中心环。Ts-serpin-1基因在猪带绦虫成虫和中绦期幼虫均表达,且成虫表达量显著高于幼虫。重组蛋白Ts-serpin-1的分子质量约为43 ku,可抑制THP-1细胞促炎性细胞因子IL-6、IL-1β、IL-12、TNF-α、IFN-γ和iNOS2的表达,促进抗炎性细胞因子IL-10和TGF-β的分泌表达。以上结果提示,Ts-serpin-1在寄生虫-宿主互作过程中可能发挥重要作用。A serine protease inhibitor Ts-serpin-1(WormBase:TSM000065700)of Taenia solium was investigated for the immunomodulatory effects on THP-1 cells.The coding DNA sequence(CDS)of Ts-serpin-1 was amplified by RT-PCR with specific primers.qRT-PCR was used to analyze the expression of Ts-serpin-1 gene in adult and larval stages.The recombinant protein Ts-serpin-1 was expressed and purified based on prokaryotic expression vector pColdⅠ.After treatment with recombinant protein Ts-serpin-1,the changes of cytokines in THP-1 cells were detected by qRT-PCR and ELISA.The sequence analysis showed that Ts-serpin-1 CDS was 1149 bp in length,encoding a protein with 382 amino acids,which contains a specific reactive centre loop motif.The results of qRT-PCR revealed that expression of Ts-serpin-1 significantly higher in adult stage than that in larval stage.The recombinant protein Ts-ser pin-1 was expressed as a fusion with a molecular weight of 43 ku.The recombinant protein Ts-serpin-1 decreased the expression of pro-inflammatory cytokines IL-6,IL-1β,IL-12,TNF-α,IFN-γand iNOS2 in the THP-1 cells that were stimulated by the recombinant protein and LPS and promoted the secretion and expression of anti-inflammatory cytokines IL-10 and TGF-βin THP-1 cells stimulated by IL-4.These results suggest that Ts-serpin-1 may play an important role in parasite-host interactions.

关 键 词：猪带绦虫 丝氨酸蛋白酶抑制剂 细胞因子 免疫调节 

分 类 号：S852.734[农业科学—基础兽医学]