细粒棘球绦虫重组Bb-Eg95-EgA31疫苗诱导小鼠免疫应答的动态观察

Dynamic observation on immune responses in mice administrated with a recombinant Bb-Eg95-EgA31 vaccine of Echinococcus granulosus

目的:观察细粒棘球绦虫(Eg)重组双歧杆菌(Bb)-Eg95-EgA31疫苗免疫小鼠后其免疫应答的动态变化。方法:将疫苗分别采用口服灌胃和鼻腔内接种免疫BALB/c鼠,分别于免疫后2、4、6、8、10、12、14、16、18和20周用ELISA法测定免疫小鼠血清中IgG及其亚类和IgE水平。用MTT法测定脾淋巴细胞的增殖,用ELISA法检测脾淋巴细胞培养上清液中IL-12、IFN-γ、TNF-α和IL-10水平,用流式细胞术(FCM)检测脾CD4+和CD8+T细胞百分率。结果:口服免疫小鼠血清IgG、IgG2a、IgG2b、IgG1、IgG3和IgE水平分别在免疫后8、2、6、6、8和10周达到峰值。脾淋巴细胞悬液中IL-12、IFN-γ、TNF-α和IL-10水平分别在免疫后4、2、4和6周达到峰值。脾淋巴细胞增殖在免疫后6周达到峰值;脾CD4+T细胞在免疫后6周达到峰值,脾CD8+T细胞无明显变化。鼻腔内接种免疫小鼠血清IgG、IgG2a、IgG2b、IgG1、IgG3和IgE水平分别在免疫后10、6、10、8、8和10周达到峰值。脾淋巴细胞悬液中IL-12、IFN-γ、TNF-α和IL-10水平分别在免疫后2、2、4和8周达到峰值。脾淋巴细胞增殖在免疫后6周达到峰值;脾CD4+T细胞在免疫后6周达到峰值,脾CD8+T细胞无明显变化。口服免疫和鼻腔内接种是两种较好的免疫途径,且前者优于后者。结论:细粒棘球绦虫重组Bb-Eg95-EgA31疫苗可诱导小鼠产生有效的免疫应答。

AIM：To dynamically observe the immune responses in mice administrated with a recombinant Bifidobacteria bifidum（Bb）-Eg95-EgA31 vaccine of Echinococcus granulosus.METHODS：BALB/c mice were immunized orally or intranasally by the vaccine.At indicated times after vaccination,the levels of serum IgG,IgG subclass and IgE were determined by ELISA.The levels of IL-12,IFN-γ,TNF-α and IL-10 were measured by ELISA in the supernatant of cultured splenocytes.Proliferation of splenocytes was tested by MTT.The percentage of spleen CD4＋ and CD8＋ cells was determined by flow cytometry（FCM）.RESULTS：The orally immunized mice achieved IgG and IgG3 peaks on 8th week post vaccination,IgG2a peak on 2th week,IgG2b and IgG1 peaks on 6th week,and IgE peak on 10th week.IFN-γ and TNF-α reached highest level on 4th week,IL-12 on 2th week,and IL-10 on 6th week.Splenocytes proliferated fastest on 6th week.The percentage of CD4＋ cells was peaked on 6th week,while the percentage of CD8＋ cells had no obvious change.The intranasally immunized group achieved peaks of IgG,IgG2b and IgE on 10th week,IgG2a peak on 6th week,and peaks of IgG1 and IgG3 on 8th week.IFN-γ and IL-12 reached highest level on 2th week,TNF-α on 4th week,and IL-10 on 8th week.Splenocytes proliferated fastest on 6th week.The percentage of CD4＋ cells was peaked on 6th week,while the percentage of CD8＋ cells had no obvious change.CONCLUSION：The recombinant Bb-Eg95-EgA31 vaccine of Echinococcus granulosus induced effective immune responses in mice by either oral vaccination or intranasal inoculation.Our data showed the better immunogenicity of oral vaccination.