目的 探讨以表面展示免疫球蛋白结合蛋白功能域(FDIBP)的枯草芽孢杆菌芽孢制备的黏膜佐剂不同途径免疫的应用效果.方法 分别将表面展示FDIBP的重组枯草芽孢杆菌芽孢及大肠杆菌不耐热毒素(LT)与人IgG(hIgG)作为佐剂共孵育制备疫苗,通过灌胃、滴鼻途径免疫BALB/c小鼠,5周后收集外周血、肺泡、小肠及阴道冲洗液.采用ELISA方法检测血清中IL-2、IL-4、IL-5、IFN-γ抗hIgG特异性IgG抗体,同时检测血清及肺泡、小肠、阴道冲洗液中抗hIgG特异性IgA抗体水平.结果 与无佐剂免疫者比较,枯草芽孢杆菌芽孢佐剂滴鼻能有效提高小鼠体液免疫产生的抗原特异性抗体IgA及IgG水平(P＜0.01),但未能有效刺激细胞免疫上调血清IL-2、IL-4、IL-5、IFN-γ水平;通过灌胃途径免疫者上述特异性抗体及细胞因子水平均无显著变化(P均＞0.05).结论 展示FDIBP的枯草芽孢杆菌芽孢可用作滴鼻途径黏膜疫苗佐剂,能有效刺激机体产生特异性黏膜免疫反应.%Objective To study the application effect of bacillus subtilis spores which displayed functional domains of immunoglobulin binding protein(FDIBP) on surface as mucosal adjuvant in different immunization routes.Methods Bacillus subtilis spores displaying FDIBP,Escherichia coli heat-labile enterotoxin (LT) and hIgG as adjuvants were incubated for preparing the vaccines.Then the BALB/c mice were separately conducted by intragastric administration and intranasal immunization.5 weeks later,the eyeballs were picked out to get blood.The mice were killed by cervical dislocation to collect flushing fluids of pulmonary alveolar,small intestine and vagina.Then the level of hIgG-specific antibody IgA in serum,lung,intestine and vagina flushing fluids and the levels of hIgG-specific antibody IgG,IL-2,IL-4,IL-5 and IFN-γ were detected by ELISA.Results Compared with those without adjuvants,bacillus subtilis spores adjuvants through intranasal immunization effectively enhanced the immunity of mice to produce antigen-specific antibodies IgA and IgG (P ＜0.01),but failed to effectively up-regulate serum levels of IL-2,IL-4,IL-5 and IFN-γ; there were no significant changes of the specific immunity antibody and cytokines in intragastric administration (all P ＞ 0.05).Conclusion The spores displaying FDIBP can be used as adjuvant in intranasal immunization and effectively stimulate the body to produce special mucosal immune response.
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