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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。
OBJECTIVE To enhance the immunogenicity of peptide drugs, the recombinant PEB1-LTB polyacrylic acid resin microspheres with fusion protein PEB1 and Escherichia coli heat-labile enterotoxin B subunit (LTB) were prepared and analyzed. In vivo humoral immune responses and cellular immune response levels in Balb / c mice immunized. Methods The microspheres were prepared by emulsion - solvent evaporation method. The microsphere size, entrapment efficiency and in vitro release were measured. Female Balb / c mice were randomly divided into 4 groups and immunized on the 0th, the 14th and the 30th day respectively. After the last immunization for 14 days, serum specific IgG, IgA, sIgA in mucosal washings and spleen cell culture The supernatant IFN-γ, IL-4 content. Results Polyacrylic acid resin microspheres with the average particle size of (47.6 ± 10.2) μm were prepared. The encapsulation efficiency was 79.87%. The microspheres were able to tolerate the destruction of gastric acid, which reached 95.27% after 4 h in simulated intestinal fluid. After immunization, the specific IgG, IgA, sIgA, IFN-γ and IL-4 were significantly higher than those in the control group (P <0.05). Conclusion The recombinant PEB1-LTB polyacrylate resin microspheres prepared in this study can effectively enhance its immune response by oral immunization of Balb / c mice.