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
发布时间：2018-11-29 12:06
【摘要】：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。
[Abstract]:Objective to investigate the effects of different adjuvants and immune pathways on the immunogenicity of Mycobacterium tuberculosis multiepitope fusion protein (TP15). Methods the multiepitope fusion protein (TP15,) was prepared using MF59 and h BCG as adjuvants, respectively, and were injected subcutaneously or intramuscularly into BALB/c mice. The serum anti-TP15 specific Ig G g G 1 and Ig G 2a antibodies were detected by indirect ELISA assay. The content of multifunctional CD4~ T cells in spleen lymphocytes and the levels of cytokines IL-1 尾 and IL-12 in the supernatant of murine peritoneal macrophages were detected by flow cytometry and sandwich ELISA. Results 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 were used as adjuvant of TP15 antigen alone or in combination. Both subcutaneous injection and intramuscular injection could significantly increase the levels of antibodies against TP15 specific Ig G g G 1 and Ig G 2a, and the immunogenicity of complex adjuvant MF59 h BCG was better. MF59 and h BCG were used as TP15 antigen adjuvant alone or in combination. Subcutaneous injection was more favorable than intramuscular injection in producing Ig G2a antibody, and the mice immunized with MF59 h BCG vaccine with complex adjuvant had the best effect on producing G2a antibody. The value of Ig G1/Ig G2a was significantly lower than that of intramuscular injection. MF59 and h BCG were immunized subcutaneously with TP15 antigen adjuvant, the content of IL-2~ CD4~ T cells and IL-10~ CD4~ T cells in spleen lymphocytes increased, and IL-10~ CD4~ T cells decreased. The levels of IL-1 尾 and IL-12 were enhanced by peritoneal macrophages. Conclusion MF59 h BCG combined adjuvant induces Th1 type cell immune drift, and the immune effect of subcutaneous injection is better than that of intramuscular injection.
【作者单位】： 解放军第三0九医院器官移植与免疫调节北京市重点实验室;西南大学生命科学学院;
【基金】：国家重大传染病专项(2013ZX10003006-003-001)
【分类号】：R392
本文编号：2364950
[Abstract]:Objective to investigate the effects of different adjuvants and immune pathways on the immunogenicity of Mycobacterium tuberculosis multiepitope fusion protein (TP15). Methods the multiepitope fusion protein (TP15,) was prepared using MF59 and h BCG as adjuvants, respectively, and were injected subcutaneously or intramuscularly into BALB/c mice. The serum anti-TP15 specific Ig G g G 1 and Ig G 2a antibodies were detected by indirect ELISA assay. The content of multifunctional CD4~ T cells in spleen lymphocytes and the levels of cytokines IL-1 尾 and IL-12 in the supernatant of murine peritoneal macrophages were detected by flow cytometry and sandwich ELISA. Results 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 were used as adjuvant of TP15 antigen alone or in combination. Both subcutaneous injection and intramuscular injection could significantly increase the levels of antibodies against TP15 specific Ig G g G 1 and Ig G 2a, and the immunogenicity of complex adjuvant MF59 h BCG was better. MF59 and h BCG were used as TP15 antigen adjuvant alone or in combination. Subcutaneous injection was more favorable than intramuscular injection in producing Ig G2a antibody, and the mice immunized with MF59 h BCG vaccine with complex adjuvant had the best effect on producing G2a antibody. The value of Ig G1/Ig G2a was significantly lower than that of intramuscular injection. MF59 and h BCG were immunized subcutaneously with TP15 antigen adjuvant, the content of IL-2~ CD4~ T cells and IL-10~ CD4~ T cells in spleen lymphocytes increased, and IL-10~ CD4~ T cells decreased. The levels of IL-1 尾 and IL-12 were enhanced by peritoneal macrophages. Conclusion MF59 h BCG combined adjuvant induces Th1 type cell immune drift, and the immune effect of subcutaneous injection is better than that of intramuscular injection.
【作者单位】： 解放军第三0九医院器官移植与免疫调节北京市重点实验室;西南大学生命科学学院;
【基金】：国家重大传染病专项(2013ZX10003006-003-001)
【分类号】：R392
【共引文献】
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1 宋庆德;黄香玉;朱传智;李军丽;肖漓;高钰;马锡慧;孔祥瑞;何秀云;;MF59/hBCG复合佐剂促进巨噬细胞募集和分泌MCP-1[J];免疫学杂志;2015年08期
，本文编号：2364950
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