中华鲟IFN-γ的免疫调控作用  

Immune regulation role of IFN-γin Acipenser sinensis

作　　者：郑楚文 袁汉文[1] 田光明[1] 李骞 张书环[2] 许巧情[1] 危起伟[2] ZHENG Chuwen;YUAN Hanwen;TIAN Guangming;LI Qian;ZHANG Shuhuan;XU Qiaoqing;WEI Qiwei(School of Animal Science,Yangtze University,Jingzhou 434020,China;Key Lab of Freshwater Biodiversity Conservation Ministry of Agriculture and Rural Affairs,Yangtze River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Wuhan 430223,China)

机构地区：[1]长江大学动物科学学院,湖北荆州434020 [2]中国水产科学研究院长江水产研究所,农业农村部淡水生物多样性保护重点实验室,湖北武汉430223

出　　处：《水产学报》2020年第9期1539-1548,共10页Journal of Fisheries of China

基　　金：农业农村部淡水生物多样性保护重点实验室开放基金(LFBC 1007)。

摘　　要：为了解中华鲟IFN-γ的免疫调控作用,实验通过中华鲟转录组获得IFN-γcDNA序列,构建重组表达质粒pTRI-st-IFNγ,原核表达IFN-γ蛋白,检测其在病毒和细菌感染过程中的免疫调控作用。SDS-PAGE显示,中华鲟IFN-γ重组蛋白大小为19.17 ku,以可溶性表达为主,浓度为0.998 mg/mL;荧光定量PCR显示,IFN-γ蛋白能够诱导中华鲟肾脏细胞中下游相关抗病毒基因Mx、Viperin和CXCL家族中CXCL11-L1、CXCL11-L2表达;与EPC细胞共孵育能够有效抑制鲤春病毒血症病毒(spring viraemia of carp virus,SVCV)引起的细胞病变和增殖,并对SVCV病毒的G、P、N 3个基因表达量呈现出极显著性下调;在体外能够调控抑制嗜水气单胞菌、中间产气单胞菌和维氏气单胞菌并对其生长表现出明显的抑制作用。Acipenser sinensis belongs to the cartilaginous fish,which is between the cartilaginous fishes and teleost fishes.It is an important branch point in the evolutionary tree of the biological immune system.It is a living fossil for the study of the evolution of fish and vertebrates,and plays an extremely important role in the evolutionary history of fish and vertebrates.In order to understand the immunomodulatory effect of A.sinensis IFN-γ,IFN-γsequence was obtained by transcriptome of A.sinensis,the recombinant plasmid of pTRI-st-IFNg was constructed and transformed into the BL21 Escherichia coli for expression and expressed in competent prokaryotic cells.SDS-PAGE analysis showed that the size of IFN-γrecombinant protein was 19.17 ku,and the concentration was 0.998 mg/mL.Quantitative PCR showed that the antiviral genes Mx,Viperin,CXCL11-L1,CXCL11-L2(CXC subfamily of chemokines)and antiviral protein Epithelioma papulosum cyprini cells(EPC)P,N,G were induced,which indicated that IFN-λactivated antiviral activity.Additionally,spring viremia of carp virus(SVCV)replication as well as the production of cytopathic effect(CPE)was significantly inhibited.Results of incubation with Aeromonas hydrophila,A.intermedia and A.veronii showed a significant inhibition on their growth and activity.In conclusion,this study provides partial theory for the understanding of IFN-γbioactivity in A.sinensis.

关 键 词：中华鲟 IFN-Γ 免疫调控 生物学功能 

分 类 号：S941[农业科学—水产养殖]