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目的探讨α-甘露糖肽(α-mannatide)作为佐剂对恶性疟原虫多表位蛋白疫苗M.RCAg-1免疫原性的影响。方法将小鼠随机分为4组:疫苗组(M.RCAg-1)、疫苗佐剂组(M.RCAg-1+α-mannatide)、佐剂对照组(α-mannatide)、弗氏佐剂对照组(M.RCAg-1+Freund’s),分别于第0、14、28天经背部皮下免疫小鼠,0.1 ml/只。采用ELISA法检测3次免疫后小鼠血清Ig G抗体水平、抗体亚型以及血清抗体对单表位的识别,间接免疫荧光反应(IFA)检测抗体对恶性疟原虫天然蛋白的识别,ELISPOT检测其细胞因子的分泌水平。结果 3次免疫后,疫苗佐剂组GMT明显高于疫苗组(P<0.05);疫苗佐剂组GMT与弗氏佐剂对照组持平(P>0.05)。疫苗佐剂组产生的高水平Ig G主要以Ig G1为主。疫苗佐剂组抗体对单表位和天然虫体的识别效果显著。免疫小鼠各表位和蛋白诱发分泌IL-4的特异性淋巴细胞克隆数高于分泌IFNγ的特异性淋巴细胞克隆数(P<0.05)。结论α-mannatide作为恶性疟原虫多表位蛋白疫苗M.RCAg-1的佐剂能诱导其产生高水平的体液免疫和细胞免疫应答,具有一定的抗疟原虫感染保护作用。
Objective To investigate the effect of α-mannatide as adjuvant on the immunogenicity of M.RCAg-1 Plasmodium falciparum multi-epitope protein vaccine. Methods Mice were randomly divided into 4 groups: vaccine group (M.RCAg-1), vaccine group (M.RCAg-1 + α-mannatide), adjuvant control group (α-mannatide), Freund’s adjuvant In the control group (M.RCAg-1 + Freund’s), mice were immunized subcutaneously on the 0th, 14th and 28th day respectively with 0.1 ml / mouse. ELISA was used to detect the level of Ig G antibody in serum, antibody subtype and serum antibody to single epitopes after three immunizations. Indirect immunofluorescence reaction (IFA) was used to detect the antibody’s recognition of P. falciparum natural protein. ELISPOT The level of cytokine secretion. Results After three immunizations, the GMT of the vaccine adjuvant group was significantly higher than that of the vaccine group (P <0.05). The GMT of vaccine adjuvant group was the same as that of the Freund adjuvant control group (P> 0.05). The high level of Ig G produced by the vaccine adjuvant group was dominated by Ig G1. Vaccine adjuvant antibody group of single epitopes and natural recognition of significant effect. The number of specific lymphocytes secreting IL-4 secreted by each epitope and protein of immunized mice was higher than the number of specific lymphocytes secreting IFNγ (P <0.05). Conclusion α-mannatide can induce high level of humoral and cellular immune response as an adjuvant to Plasmodium falciparum polyepitope vaccine M.RCAg-1, and has certain protection against Plasmodium infection.