H9亚型禽流感病毒M1蛋白原核表达及免疫效果初步研究  

Preliminary Study on the Immune Effect of H9 Subtype AIV M1 Protein by Prokaryotic Expression

作　　者：李小凤 谢芝勋 阮志华 李孟 李丹 张民秀 谢志勤 罗思思 韦悠 谢丽基 曾婷婷 张艳芳 黄娇玲 王盛 LI Xiao-feng;XIE Zhi-xun;RUAN Zhi-hua;LI Meng;LI Dan;ZHANG Min-xiu;XIE Zhi-qin;LUO Si-si;WEI You;XIE Li-ji;ZENG Ting-ting;ZHANG Yan-fang;HUANG Jiao-ling;WANG Sheng(Guangxi Key Laboratory of Veterinary Biotechnology,Key Laboratory of China(Guangxi)-ASEAN Cross-border Animal Disease Prevention and Control,Ministry of Agriculture and Rural Affairs of China,Guangxi Veterinary Research Institute,Nanning,Guangxi,530001,China)

机构地区：[1]广西壮族自治区兽医研究所/广西兽医生物技术重点实验室/农业农村部中国(广西)-东盟跨境动物疫病防控重点实验室,广西南宁530001

出　　处：《动物医学进展》2024年第6期1-6,共6页Progress In Veterinary Medicine

基　　金：广西重点研发计划项目(桂科AB21076004);广西自然科学基金项目(2022GXNSFAA035445,2021GXNSFBA196031);广西八桂学者项目(2019-A50)。

摘　　要：旨在原核表达H9亚型禽流感病毒(AIV)M1蛋白,初步探究其免疫效果,为研制AIV疫苗奠定基础。通过构建重组质粒pET-32a-M1,诱导表达,Western blot验证蛋白表达情况,将重组蛋白免疫接种小鼠,通过测定血清中IgG抗体效价,脾细胞培养上清多细胞因子分泌情况评价其免疫效果。结果显示,M1重组蛋白主要以可溶性蛋白存在,Western blot结果可见特异性条带。免疫接种小鼠后,血清产生IgG抗体,抗体效价为1∶30000。小鼠脾细胞上清液IL-2、IL-4、IL-6与对照组相比较,含量显著增加(P<0.01),而抑制IL-1β、IL-5、IL-13、IL-12p70、IL-18、GM-CMF、IFN-γ、TNF-α分泌。结果表明,M1重组蛋白免疫原性良好,刺激机体产生体液免疫和细胞免疫,为禽流感疫苗深入研究奠定了基础。This study aims to express the M1 protein of the H9 subtype avian influenza virus(AIV)and preliminarily explore its immune effect,laying the foundation for the development of avian influenza virus(AIV)vaccine.By constructing the recombinant plasmid pET-32a-M1,inducing and Western-blot to verify protein expression,immune mice with recombinant proteins,and the immune effect of IgG antibody titer in serum and supernatant multicytokine secretion in splenocytes culture was evaluated.The results showed that the expressed protein existed in soluble forms,Western-blot results showed specific bands.Serum of immunized mice produced IgG antibody,antibody titer was 1∶30000.Compared with the control group,the contents of supernatant IL-2,IL-4,IL-6 in mouse spleen cells were significantly increased,respectively(P<0.01).It inhibits IL-1β,IL-5,IL-13,IL-12p70,IL-18,GM-CMF,IFN-γ,TNF-α.Conclusion,the prokaryotically expressed M1 protein had good immunogenicity,can stimulate humoral immunity and cellular immunity,laying foundation for further research on influenza vaccine.

关 键 词：禽流感 M1蛋白 原核表达 免疫效果 

分 类 号：S852.659.5[农业科学—基础兽医学] Q789[农业科学—兽医学]