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目的研究日本血吸虫重组双歧杆菌属两歧双歧杆菌(Bb)(pGEX-Sj14-3-3)疫苗免疫小鼠后其免疫应答的动态变化。方法将重组疫苗分别采用口服灌胃和鼻内接种免疫BALB/c小鼠,分别于免疫后2、4、6、8、10、12、14、16、18、20和22周ELISA法测小鼠血清中IgG及其亚类、IgE和IgA及脾细胞培养液中IFN-γ、IL-12、TNF-α和IL-10水平;MTT法测定脾细胞增殖;流式细胞仪检测脾细胞凋亡率及CD4+和CD8+T细胞亚群百分率。结果口服组血清IgG、IgG1、IgG2a、IgG2b、IgG3、IgE和IgA水平分别在免疫后8、6、6、4、8、10和6周达峰值。脾淋巴细胞增殖和凋亡水平均在免疫后4周达峰值;CD4+T细胞于8周达峰值,CD8+T细胞于14周达峰值。IFN-γ、IL-12、TNF-α和IL-10水平分别于8、8、6和4周达峰值。鼻内接种组血清IgG、IgG1、IgG2a、IgG2b、IgG3、IgE和IgA分别于4、6、4、4、8、10和8周达峰值。脾淋巴细胞增殖及凋亡水平也均在4周达峰值;CD4+T细胞于8周达峰值,CD8+T细胞于4周达峰值。IFN-γ、IL-12、TNF-α和IL-10水平分别于2、2、4和4周达峰值。口服及鼻腔内接种是两种较好的免疫途径,且后者优于前者。结论该疫苗可诱导小鼠产生有效的免疫应答。
Objective To study the dynamic changes of immune response in mice immunized with recombinant Bifidobacterium spp. Bifidobacterium japonicum (pGEX-Sj14-3-3) vaccine. Methods BALB / c mice were immunized with recombinant vaccine by oral gavage and intranasal inoculation, respectively, and were detected by ELISA at 2,4,6,8,10,12,14,16,18,20 and 22 weeks after immunization The levels of IFN-γ, IL-12, TNF-α and IL-10 in IgG and its subclasses, IgE and IgA in rat serum and splenocytes culture medium were measured. The proliferation of spleen cells was determined by MTT assay. Mortality and percentage of CD4 + and CD8 + T cell subsets. Results Serum levels of IgG, IgG1, IgG2a, IgG2b, IgG3, IgE and IgA in oral group reached the peak at 8th, 6th, 6th, 4th, 4th, 8th, 10th, and 6th week after immunization. Splenic lymphocyte proliferation and apoptosis levels peaked at 4 weeks after immunization; CD4 + T cells peaked at 8 weeks and CD8 + T cells reached their peak at 14 weeks. The levels of IFN-γ, IL-12, TNF-α and IL-10 peaked at 8, 8, 6 and 4 weeks respectively. Serum IgG, IgG1, IgG2a, IgG2b, IgG3, IgE and IgA peaked at 4, 6, 4, 4, 8, 10 and 8 weeks respectively in the intranasal vaccination group. The proliferation and apoptosis of splenic lymphocytes also peaked at 4 weeks, CD4 + T cells peaked at 8 weeks and CD8 + T cells reached peak at 4 weeks. The levels of IFN-γ, IL-12, TNF-α and IL-10 peaked at 2, 2, 4 and 4 weeks respectively. Oral and intranasal vaccination are two better routes of immunization, and the latter is better than the former. Conclusion The vaccine can induce mice to produce an effective immune response.