表达猪流行性腹泻病毒中和抗原位点的重组干酪乳杆菌免疫小鼠诱导的免疫应答  

Immune response in mice after oral inoculation with Lactobacillus casei expressing the neutralizing epitope of spike protein of porcine epidemic diarrhea virus

作　　者：葛俊伟[1] 姜艳平[1] 汪淼[1] 乔薪瑗[1] 刘敏[1] 唐丽杰[1] 李一经[1] 

机构地区：[1]东北农业大学动物医学院,黑龙江哈尔滨150030

出　　处：《中国兽医科学》2010年第7期708-712,共5页Chinese Veterinary Science

基　　金：黑龙江省"十一五"科技攻关项目(GA06B202-4);东北农业大学博士启动基金项目(2009)

摘　　要：为研究猪流行性腹泻病毒中和抗原位点(COE)基因在乳酸菌中的表达情况,进而探讨重组乳酸菌的免疫原性。将COE基因插入干酪乳杆菌细胞表面表达载体pPG-1中,构建重组表达载体pPG-1-COE,并将其电转化入干酪乳杆菌Lactobacilluscasei393中,应用Western-blotting、间接免疫荧光染色和全细胞ELISA检测重组蛋白的表达情况,用重组干酪乳杆菌口服免疫小鼠,测定免疫应答水平。结果表明,COE可在构建的重组干酪乳杆菌表达系统中表达,目的蛋白定位于细胞表面。在免疫组小鼠血清、粪便中检测到较高水平的特异性IgG、sIgA(P<0.01);血清具有中和活性(1∶12);脾淋巴细胞增殖指数明显高于对照组,IL-4和IFN-γ水平显著提高(P<0.01)。证实,该重组干酪乳杆菌表达系统可诱导小鼠产生对猪流行性腹泻病毒特异的黏膜免疫应答和系统免疫应答。To evaluate the immunogenicity of recombinant Lactobacillus casei 393 expressing the neutralizing epitope（COE）of porcine epidemic diarrhea virus（PEDV）,a recombinant Lactobacillus expressing COE was constructed with plasmid pPG-1,and expression and localization of the COE protein from recombinant pPG-1-COE/L.casei 393 were examined by Western-blotting,indirect immunofluorescence and the whole bacteria ELISA.The results showed that the recombinant COE protein displayed antigenic specificity and was located on the bacterial cell walls.Both local mucosal and systemic immune responses against PEDV were induced in the BALB/c mice immunized orally with the recombinant Lactobacillus.The induced antibodies demonstrated neutralizing effects on PEDV infection.Specific cellular immune response was also observed.In conclusion,the oral immunizations with recombinant L.casei 393 could induce significant specific mucosal PEDV specific IgA response as well as serum IgG responses,and could evoke both mucosal and systemic immune responses.

关 键 词：猪流行性腹泻病毒 中和抗原位点 干酪乳杆菌 免疫反应 

分 类 号：S852.659.6[农业科学—基础兽医学]