注射用布氏菌活疫苗免疫学评价

目的以小鼠为动物模型,通过检测布氏菌活疫苗免疫小鼠产生的体液免疫、细胞免疫应答及保护力,对其进行免疫学评价。方法将30只小鼠随机分为3组,皮下免疫布氏活疫苗,免疫4周后分离血清及脾脏淋巴细胞。ELISA法检测免疫动物血清中总IgG抗体,抗体亚类IgG1及IgG2a;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IFN-γ、IL-4的细胞因子水平;用羊布氏菌弱毒株M5攻击免疫动物,通过脾脏细菌计数评价布氏活疫苗的免疫保护作用。结果用ELISA法检测到免疫布氏活苗的小鼠体内有特异性抗体产生;ELISPOT可检测到较高的分泌IFN-γ脾脏淋巴细胞数目,ELISA方法可检测到较高的IFN-γ水平,表明布氏活疫苗主要诱导Th1型免疫应答;通过布氏活疫苗保护力分析,表明该疫苗具有较好的免疫保护效果。结论布氏疫苗采用注射免疫途径是可行的,免疫后能获得较好的免疫效果。

Evalution of immunogical response of live Brucella vaccine for injection

Objective To evaluate the immune response of live Brucella vaccine for intradermal injection, which detect humoral immunity and cellular immune responses in mice. Methods Mice were randomly divided into 3 groups with intradermal immune live Brucella vaccine, After 4 weeks, blood was sampled and spleen lymphocytes were separated. Then serum ELISA was used to detect the immune animal total IgG antibody, IgG1, IgG2a The number of spleen lymphocytes with secretion of IFN-γ, IL-4 was examined by ELISPOT. ELISA method was adopted to detect the level of cytokines IFN-γ, IL-4 from spleen cells after restimulation in vitro. After Brucella melitensis M5 infected animals, the spleen were isolated in sterile. The acquired immune protection of immune Brucella vaccine was evaluated through the bacteria count of spleen. Results The ELISA results showed that there exerted specific antibodies in mice infected by immune live Brucella vaccine. The number of spleen lymphocytes with secretion of INF-γwas increased through ELISPOT assay. The level of IFN-γwas higher than the control after detection of ELISA method. From this, live Brucella vaccine for intradermal injection mainly induced type TH1 immune response. The results from analysis of protection force of live Brucella vaccine demonstrated that the vaccine produced strong immune protection. Conclusion The method of live Brucella vaccine for intradermal injection is feasible, which can get intensively immunogenicity.