空肠弯曲菌重组蛋白微球疫苗制备及其免疫原性的分析

Construction and immunogenicity evaluation of Campylobacter jejun fusion protein microparticles vaccines

目的为增强多肽蛋白类药物的免疫效果,制备空肠弯曲菌外膜蛋白PEB1与大肠埃希菌不耐热肠毒素B亚单位(LTB)融合表达的重组PEB1-LTB蛋白聚丙烯酸树脂微球,分析免疫Balb/c小鼠体内体液免疫应答及细胞免疫应答水平。方法乳剂—溶剂挥发法制备微球,测量微球粒径、包封率和体外释放度。雌性Balb/c小鼠随机分为4组,于第0、14、30天各免疫1次,末次免疫14 d后,检测各组血清特异性IgG、IgA、黏膜冲洗液sIgA水平及脾细胞培养上清中IFN-γ、IL-4含量。结果制备得到平均粒径在(47.6±10.2)μm的聚丙烯酸树脂微球,包封率为79.87%。微球可耐受胃酸的破坏,在模拟肠液中的释放度4 h后达95.27%。免疫后特异性IgG、IgA、sIgA及IFN-γ、IL-4均明显高于对照组(P<0.05)。结论本研究制备的重组PEB1-LTB蛋白聚丙烯酸树脂微球通过口服免疫Balb/c小鼠可以有效的提高其免疫应答。

To improve the immunogenicity of protein vaccine, we prepared Campylobacter jejuni recombinant PEB1-LTB protein polyacrylic resin microsphere, and validate it in Balb/c mice. By oil-in-oil emulsion solvent evaporation method, the microparticle was prepared with particle size of（47.6±10.2） μm, entrapment efficiency of79.87%, and accumulative release rate of 95.27% in mimic intestinal fluid of 4 h. Then the microparticle was used to immunize Balb/c on days 0, 14, 30. Fourteen days after last immunization, the levels of IgG, IgA and sIgA antibody levels, as well as the IFN-γ and IL-4 levels in splenocyte culture supernatant of mice were detected. Data showed that the levels of IgG, IgA, sIgA, IFN-γ, and IL-4 were increased obviously in the microparticle immunized mice, as compared with control groups（P〈0.05）. In conclusion, the Campylobacter jejuni recombinant PEB1-LTB protein polyacrylic resin microsphere prepared in this study can improve the immunity of mice.