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利用非复制型痘苗病毒表达载体 pNEOCK11β75IL5和重组病毒RVJ12 3,通过两步重组构建了能同时表达IL 5和乙型肝炎病毒HBsAg的非复制型重组痘苗病毒RVJ12 3Δ11β75IL5。Southern blot证实 ,痘苗病毒C K片段间基因缺失的同时伴有IL 5基因的插入。鼻腔吸入分别免疫Balb/c小鼠和新西兰白兔 ,ELISPOT实验证实 ,免疫后两周小鼠肺淋巴细胞的抗HBsAgIgA抗体分泌细胞 (ASC)数比对照组 (RVJ12 3Δ11β75 )增加约 2倍 ,而同时小鼠肺淋巴细胞的抗HBsAgIgG抗体分泌细胞 (ASC)数与对照组无差别。可在小鼠血液、肺浸出液以及新西兰白兔血液、肺浸出液、其它分泌液样品中检测到抗乙型肝炎病毒HBsAg的特异性的IgA、IgG抗体 ,与对照组相比 ,IgA抗体阳转率及抗体滴度提高 ,而IgG则无差异。本实验说明 :IL 5可在体内选择性地增强机体的粘膜IgA反应。提示非复制载体疫苗中 ,表达的该细胞因子可有效的增强疫苗的粘膜免疫反应 ,为粘膜疫苗的发展策略提供了新的途径
A non-replicating recombinant vaccinia virus RVJ12 3Δ11β75IL5 was constructed by two-step recombination using the non-replicating vaccinia virus expression vector pNEOCK11β75IL5 and recombinant virus RVJ12 3, which can express both IL-5 and hepatitis B virus HBsAg simultaneously. Southern blot confirmed that the deletion of the gene between the vaccinia C fragment and IL 5 gene insertion. Nasal inhalation were immunized Balb / c mice and New Zealand white rabbits, ELISPOT experiments confirmed that the number of anti-HBsAgIgA antibody secreting cells (ASC) of lung lymphocytes in mice two weeks after immunization than the control group (RVJ12 3Δ11β75) increased about 2 times At the same time, the number of anti-HBsAg IgG antibody secreting cells (ASC) in mouse lung lymphocytes did not differ from the control group. Anti-HBsAg-specific IgA and IgG antibodies were detected in mouse blood, lung leachate and New Zealand white rabbit blood, lung leachate, and other exudate samples. Compared with the control group, the IgA antibody positive rate And antibody titer increased, while there was no difference in IgG. This experiment shows that: IL 5 can selectively enhance the body’s mucosal IgA response in vivo. It is suggested that this cytokine expressed in non-replicating vector vaccine can effectively enhance mucosal immune response of vaccine and provide a new way for the development of mucosal vaccine