【摘要】：目的探討不同佐劑和免疫途徑對結核分枝桿菌多表位融合蛋白TP15免疫原性的影響。方法制備多表位融合蛋白TP15,分別以MF59和h BCG單獨或聯(lián)合作為佐劑,經皮下或肌肉注射BALB/c小鼠,采用間接ELISA法檢測小鼠血清抗TP15特異性Ig G、Ig G1和Ig G2a抗體,流式細胞術檢測小鼠脾淋巴細胞中多功能CD4~+T細胞含量,夾心ELISA法檢測小鼠腹腔巨噬細胞培養(yǎng)上清中細胞因子IL-1β和IL-12的含量。結果多表位融合蛋白TP15免疫小鼠只產生抗TP15特異性Ig G和Ig G1抗體,不產生Ig G2a抗體。MF59和h BCG單獨或聯(lián)合作為TP15抗原佐劑,皮下注射和肌肉注射均能顯著提高機體抗TP15特異性Ig G、Ig G1和Ig G2a抗體水平,且復合佐劑MF59+h BCG的免疫效果更佳。MF59和h BCG單獨或聯(lián)合作為TP15抗原佐劑,皮下注射較肌肉注射更有利于產生Ig G2a抗體,其中復合佐劑MF59+h BCG疫苗免疫小鼠產生Ig G2a抗體的效果最佳,且Ig G1/Ig G2a值顯著低于肌肉注射。MF59和h BCG單獨或聯(lián)合作為TP15抗原佐劑經皮下免疫小鼠,脾淋巴細胞IL-2~+CD4~+T細胞含量增加,IL-10~+CD4~+T細胞含量減少,腹腔巨噬細胞分泌IL-1β和IL-12水平增強。結論 MF59+h BCG復合佐劑誘導Th1型細胞免疫漂移,且皮下注射免疫效果優(yōu)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.
【作者單位】： 解放軍第三0九醫(yī)院器官移植與免疫調節(jié)北京市重點實驗室;西南大學生命科學學院;
【基金】：國家重大傳染病專項(2013ZX10003006-003-001)
【分類號】：R392

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1 宋慶德;黃香玉;朱傳智;李軍麗;肖漓;高鈺;馬錫慧;孔祥瑞;何秀云;;MF59/hBCG復合佐劑促進巨噬細胞募集和分泌MCP-1[J];免疫學雜志;2015年08期

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本文編號：2364950