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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。
Objective: To observe the dynamic changes of immune response of mice immunized with Echinococcus granulosus (Eg) recombinant Bb-Eg95-EgA31 vaccine. Methods: BALB / c mice were immunized with oral gavage and intranasal inoculation respectively. Immunized mice were determined by ELISA at 2,4,6,8,10,12,14,16,18 and 20 weeks after immunization Serum IgG and its subclasses and IgE levels. The proliferation of splenic lymphocytes was measured by MTT assay. The levels of IL-12, IFN-γ, TNF-α and IL-10 in supernatant of splenic lymphocytes were measured by ELISA. The splenic CD4 + And CD8 + T cell percentage. Results: Serum IgG, IgG2a, IgG2b, IgG1, IgG3 and IgE levels in oral immunized mice peaked at 8, 2, 6, 6, 8, 10 and 10 weeks after immunization. The levels of IL-12, IFN-γ, TNF-α and IL-10 in splenic lymphocyte suspension reached the peak at 4, 2, 4 and 6 weeks after immunization. Splenic lymphocyte proliferation peaked at 6 weeks after immunization; splenic CD4 + T cells peaked at 6 weeks after immunization, and splenic CD8 + T cells showed no significant changes. Serum IgG, IgG2a, IgG2b, IgG1, IgG3 and IgE levels in the immunized mice reached the peak at 10, 6, 10, 8, 8 and 10 weeks after immunization. The levels of IL-12, IFN-γ, TNF-α and IL-10 in splenic lymphocyte suspension peaked at 2, 2, 4 and 8 weeks after immunization. Splenic lymphocyte proliferation peaked at 6 weeks after immunization; splenic CD4 + T cells peaked at 6 weeks after immunization, and splenic CD8 + T cells showed no significant changes. Oral immunization and intranasal vaccination are two better routes of immunization, and the former is better than the latter. Conclusion: Echinococcus granulosus recombinant Bb-Eg95-EgA31 vaccine can induce effective immune response in mice.