Objective To evaluate the immune effects in the mice to be immunized by theBrucella vaccine through intranasal immunization and hypodermic injection respectively. Methods Four weeks post-immunization, blood and spleen lymphocytes and the bronchoalveolar lavage fluids were collected from each animal. Serum samples and the bronchoalveolar lavage fluids were examined for total IgG/IgA antibodies using ELISA. The number of spleen lymphocytes secreting IFN-γ, IL-4 were detected by ELISPOT and ELISA method was adopted for detection of spleen cellsin vitro stimulated to secrete IFN-γ, IL-4 levels of cytokines. Immunized animals were infected with attenuatedBrucella melitensis M5 strain through two routes of hypodermic injection and intranasal attacking and then evaluated the acquired immune protection of immunized animals through the bacteria count of spleen. Results The results from ELISA, ELISPOT and classification of T lymphocyte subsets showed that both of the two methods of immunization were able to induce humoral immunity and cellular immune responses in mice. There was no significant difference between two methods. The intranasal attacking test and the hypodermic attacking test showed that theBrucella number of logarithmic value in the spleen of the intranasal immunization group, hypodermic injection group and the negative groupwere 0, 0, 6.24 ± 0.07 and 1.69 ± 0.07, 3.03 ± 0.03, 5.29 ± 0.06, respectively. Conclusions Two methods of immunization induces humoral immunity and cellular immune responses in mice. The protective effect of the intranasal immunization group is superior to that of hypodermic injection group, suggesting thatBrucella vaccine for intranasal immunization may be an effective method.%目的：将布氏减毒活疫苗通过滴鼻和皮下注射两种方式免疫小鼠，比较两种方式的免疫效果。方法采用两种方法免疫动物，免疫4周后分离血清、脾脏淋巴细胞及制备肺泡灌洗液。ELISA 法检测免疫动物血清中总 IgG抗体；ELISA法检测免疫动物肺泡灌洗液中总 IgG、IgA抗体；ELISPOT法检测分泌 IFN-γ、IL-4的脾脏淋巴细胞数目，以及用 ELISA方法检测体外再刺激后小鼠脾细胞分泌细胞因子水平；流式细胞术对 T细胞亚群进行分类；用羊布氏菌弱毒株 M5通过滴鼻和皮下注射两种方式攻击免疫动物，通过脾脏布氏菌计数评价并比较两种免疫方法的保护效果。结果 ELISA、ELISPOT和 T淋巴细胞亚群分类结果显示，两种免疫途径均能诱导体液免疫、细胞免疫应答，两者间无显著差异。BALB/c 小鼠滴鼻攻击试验结果显示滴鼻免疫组和皮下免疫组脾脏中布氏菌数量的对数值均为0，阴性组脾脏中布氏菌数量的对数值为6.24±0.07；皮下攻击试验显示滴鼻免疫组脾脏中布氏菌数量的对数值为1.69±0.07，皮下免疫组为3.03±0.03，阴性组为5.29±0.06。结论两种免疫途径均能诱导体液免疫、细胞免疫应答，滴鼻免疫组的免疫保护作用显著优于皮下注射组，提示滴鼻免疫是一种高效的免疫途径。
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