鸡传染性支气管炎病毒QX基因型S1蛋白重组乳酸菌的构建及小鼠免疫应答  

Construction of recombinant Lactococcus lactis expressing QX genotype S1 protein of avian infectious bronchitis virus and immune response in mice

作　　者：王栎婷 曹祁峰 王雪 李冰 WANG Yueting;CAO Qifeng;WANG Xue;LI Bing(College of Animal Husbandry and Veterinary Medicine,Jinzhou Medical University,Jinzhou,Liaoning121001,China;Liaoning Yikang Biological Co.,Ltd.,Liaoyang,Liaoning 111000,China)

机构地区：[1]锦州医科大学畜牧兽医学院,辽宁锦州121001 [2]辽宁益康生物股份公司,辽宁辽阳111000

出　　处：《中国兽医学报》2023年第1期49-54,60,共7页Chinese Journal of Veterinary Science

基　　金：2021年度辽宁省教育厅科学研究资金资助项目(LJKZ0800);2021年大学生创新创业训练资金资助项目(S202110160019)。

摘　　要：为探究鸡传染性支气管炎病毒(IBV)S1蛋白重组乳酸菌对小鼠免疫应答情况,本研究将IBV CH/LN/2019毒株的S1基因进行密码子优化,构建重组质粒pNZ8149-S1,电转至乳酸乳球菌NZ3900中,应用Western blot及间接免疫荧光试验评价重组乳酸乳球菌表达水平,免疫SPF小鼠,通过间接ELISA方法分析小鼠特异性抗体、CD4+、CD8+和细胞因子(IL-4和IFN-γ)分泌情况;结果显示,成功构建重组乳酸乳球菌pNZ8149-S1/NZ3900并表达目的蛋白;小鼠免疫试验结果显示,免疫组小鼠IgG抗体水平均高于pNZ8149/NZ3900、NZ3900和PBS组(P<0.05);特异性sIgA抗体水平显著高于对照组(P<0.01);此外,重组乳酸乳球菌能诱导小鼠产生较高水平的IL-4和IFN-γ(P<0.05)。结果表明,重组乳酸乳球菌pNZ8149-S1/NZ3900可以有效刺激小鼠机体产生体液免疫和细胞免疫,为IBV新型口服疫苗的研制提供了理论参考。To explore the immune response of recombinant lactic acid bacteria with S1 protein of infectious bronchitis virus in mice,the S1 gene of IBV CH/LN/2019 strain was codon optimized to construct recombinant plasmid pNZ8149-S1,which was electrotransferred into lactic acid bacteria NZ3900.The expression level of recombinant lactic acid bacteria was evaluated by Western blot and indirect immunofluorescence test,and SPF mice were immunized.Indirect ELISA was used to analyze the secretion of specific antibodies,CD4+,CD8+and cytokines(IL-4 and IFNγ)in mice.The results showed that the recombinant pNZ8149-S1/NZ3900 of Lactococcus lactis was successfully expressed and the immune test in mice showed that the level of IgG antibody in the immunized group was higher than that in the pNZ8149/NZ3900,NZ3900 and PBS groups,and the level of specific sIgA antibody was significantly higher than that in the control group.In addition,the recombinant lactic acid bacteria could induce higher levels of IL-4 and IFN-γin mice.The results showed that recombinant Lactococcus lactis pNZ8149S1/NZ3900 could effectively stimulate humoral immunity and cellular immunity in mice,which provided a theoretical reference for the development of a new oral vaccine against IBV.

关 键 词：鸡传染性支气管炎病毒 S1基因 重组乳酸乳球菌 口服免疫 免疫应答 

分 类 号：S852.65[农业科学—基础兽医学]