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目的探讨不同佐剂和免疫途径对结核分枝杆菌多表位融合蛋白TP15免疫原性的影响。方法制备多表位融合蛋白TP15,分别以MF59和h BCG单独或联合作为佐剂,经皮下或肌肉注射BALB/c小鼠,采用间接ELISA法检测小鼠血清抗TP15特异性Ig G、Ig G1和Ig G2a抗体,流式细胞术检测小鼠脾淋巴细胞中多功能CD4~+T细胞含量,夹心ELISA法检测小鼠腹腔巨噬细胞培养上清中细胞因子IL-1β和IL-12的含量。结果多表位融合蛋白TP15免疫小鼠只产生抗TP15特异性Ig G和Ig G1抗体,不产生Ig G2a抗体。MF59和h BCG单独或联合作为TP15抗原佐剂,皮下注射和肌肉注射均能显著提高机体抗TP15特异性Ig G、Ig G1和Ig G2a抗体水平,且复合佐剂MF59+h BCG的免疫效果更佳。MF59和h BCG单独或联合作为TP15抗原佐剂,皮下注射较肌肉注射更有利于产生Ig G2a抗体,其中复合佐剂MF59+h BCG疫苗免疫小鼠产生Ig G2a抗体的效果最佳,且Ig G1/Ig G2a值显著低于肌肉注射。MF59和h BCG单独或联合作为TP15抗原佐剂经皮下免疫小鼠,脾淋巴细胞IL-2~+CD4~+T细胞含量增加,IL-10~+CD4~+T细胞含量减少,腹腔巨噬细胞分泌IL-1β和IL-12水平增强。结论 MF59+h BCG复合佐剂诱导Th1型细胞免疫漂移,且皮下注射免疫效果优于肌肉注射。
Objective To investigate the effect of different adjuvants and immunization routes on the immunogenicity of Mycobacterium tuberculosis multi-epitope fusion protein TP15. Methods The multi-epitope fusion protein TP15 was prepared. BALB / c mice were injected subcutaneously or intramuscularly with MF59 and h BCG alone or in combination as the adjuvant. The serum anti-TP15 specific Ig G and Ig G1 And Ig G2a antibody. The content of multifunctional CD4 ~ + T cells in splenic lymphocytes was detected by flow cytometry. The contents of cytokines IL-1β and IL-12 in peritoneal macrophages culture supernatants were detected by sandwich ELISA . Results The multi-epitope fusion protein TP15-immunized mice only produced anti-TP15-specific Ig G and Ig G1 antibodies and did not produce Ig G2a antibodies. MF59 and h BCG alone or in combination as TP15 antigen adjuvant, subcutaneous injection and intramuscular injection can significantly enhance the body’s anti-TP15 specific Ig G, Ig G1 and Ig G2a antibody levels, and the combined adjuvant MF59 + h BCG immune effect more good. MF59 and h BCG alone or in combination as the TP15 antigen adjuvant, subcutaneous injection of intramuscular injection is more conducive to produce Ig G2a antibody, which MF59 + h BCG vaccine immunized mice Ig G2a antibody produced the best, and Ig G1 / Ig G2a values were significantly lower than the intramuscular injection. Mice were immunized subcutaneously with MF59 and h BCG alone or in combination with TP15 antigen adjuvant. The content of IL-2 ~ + CD4 ~ + T cells in splenic lymphocytes increased and the content of IL-10 ~ + CD4 ~ + T cells decreased. Cells secrete increased levels of IL-1β and IL-12. Conclusions MF59 + h BCG complex adjuvant induced Th1-type cell immune drift, and subcutaneous immunization is better than intramuscular injection.