IL-5在非复制重组痘苗病毒中的表达及其对重组病毒免疫效果的影响

Expression of IL-5 in Recombinant Non-replicating Vaccinia Virus and Its Effects on Immunogenicity

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 。

We constructed the recombinant non replicating vaccinia virus RVJ123Δ11β75IL5 using non replicating vaccinia virus vector pNEOCK11β75IL5 and replicating vaccinia virus RVJ123,which can express IL 5 and HBsAg simultaneously. By Southern blot,it was determined that the genes between vaccinia virus HindⅢ C and K fragments were deleted and IL 5 gene was integrated stably.Giving intranasal inocula of the virus to immunize Balb/c mice and New Zealand rabbits,the elevated anti HBsAg IgA antibody secreting cells(ASC)of mouse lung lymphocytes to vectors expressing IL 5 was about two times higher than the ASC induced by control virus 14 days after immunization,and they sustained for about 4 weeks.We also detected the elevation of anti HBsAg IgA antibody secreting cells in mouse serum and lung fluid,rabbit's serum,lung fluid,saliva and nasal washing samples.No enhancement of specific IgG response was detected in both animals either locally or systemically.Our results indicate that IL 5 selectively enhances the mucosal IgA response in vivo and suggest that expression of this factor in mucosal vaccine vectors may stimulate mucosal immune reactivity,giving rise a new way to improve the mucosal vaccination strategy.