复合佐剂 MF59／hBCG 对结核病蛋白疫苗免疫原性的影响

摘要

Objective To study the effects of MF59 in combination with heat-killed BCG ( hBCG) as adjuvant on the immunogenicity of Mycobacterium tuberculosis fusion protein PstS1-LEP.Methods BALB/c mice were divided into six groups from group 1 through group 6.They were immunized with PstS1-LEP+MF59 ( group 1 ) , PstS1-LEP+MF59/hBCG ( group 2 ) , PstS1-LEP+hBCG ( group 3 ) , MF59 ( group 4 ) , PstS1-LEP (group 5) and hBCG (group 6) for three times at intervals of two weeks , respectively.The mice were sac-rificed two weeks after the last immunization .The serum samples were collected for antibodies detection .The splenic lymphocytes and peritoneal macrophages were isolated and cultured with PstS 1-LEP.Indirect ELISA and sandwich ELISA were used to detect PstS 1-LEP-specific antibodies and cytokines in the supernatants of culture , respectively.Results The level of IFN-γ, IL-1β, IgG, IgG1 and IgG2a in group 1 were higher than those in groups 4, 5 and 6 (P<0.05).The level of IL-2 and IL-4 in group 1 were higher than those in groups 4 and 6 (P<0.05).The level of IFN-γ, IL-1β, IL-12, IgG, IgG1 and IgG2a in group 2 were higher than those in groups 4, 5 and 6 (P<0.05).The level of IL-2 was higher in group 2 than that in groups 4 and 6 (P<0.05). The level of IL-4 in group 3 was higher than that in group 4 ( P=0.05 ) .The level of IL-1βin group 3 were higher than that in groups 4 and 5 ( P<0.05 ) .The level of IgG was higher in group 3 than that in groups 4 and 6 (P<0.05).IgG1 level in group C was up-regulated in comparison with that in groups 4, 5 and 6 (P<0.05 ) .Conclusion hBCG as PstS1-LEP adjuvant induces a shift towards Th 2-type immune response , while MF59 induces Th1/Th2-type immune response.The combination of MF59 and hBCG inhibits the secretion of IL-4 by spleen lymphocytes , but enhances the secretion of IL-12 by macrophage .%目的：探讨复合佐剂[MF59和热灭活BCG（hBCG），MF59/hBCG]对结核分枝杆菌重组融合蛋白PstS1-LEP 免疫原性的影响。方法 BALB/c 小鼠分为6组，1组：MF59＋PstS1-LEP；2组：MF59/hBCG＋PstS1-LEP；3组：hBCG＋PstS1-LEP；4组：MF59；5组：PstS1-LEP；6组：hBCG。分别于第0、2、4周皮下免疫小鼠。末次免疫2周后解剖小鼠，PstS1-LEP体外刺激培养脾细胞和腹腔巨噬细胞。间接ELISA检测血清抗PstS1-LEP抗体，夹心ELISA检测培养上清细胞因子含量。结果1组IFN-γ、IL-1β、IgG、IgG1和IgG2a水平显著高于4组、5组和6组（P＜0.05），IL-2和IL-4水平显著高于4组和6组（P＜0.05）。2组IFN-γ、IL-1β、IL-12、IgG、IgG1和IgG2a水平显著高于4组、5组和6组（P＜0.05），IL-2水平显著高于4组和6组（ P＜0.05）。3组IL-4水平显著高于4组（ P＝0.05），IL-1β水平显著高于4组和5组（ P＜0.05），IgG水平显著高于4组和6组（ P＜0.05）， IgG1水平显著高于4组、5组和6组（ P＜0.05）。结论以PstS1-LEP为抗原，hBCG佐剂偏向诱导Th2型免疫反应，MF59佐剂诱导Th1/Th2型免疫反应，MF59和hBCG联合抑制脾细胞分泌IL-4，但增强巨噬细胞分泌IL-12。