【摘要】 目的:构建含有分子伴侣热休克蛋白70(HSP70)和甲胎蛋白(AFP)分子的蛋白复合物,研究此蛋白复合物是否具有针对AFP肿瘤诱导产生特异性CTL反应和保护性效应的能力。方法:戊二醛交联HSP70和AFP构建HSP70-AFP复合物。皮下注射免疫Balb/c纯系小鼠,同时设立单独AFP和HSP70免疫小鼠组作为对照。酶联免疫斑点实验(ELISPOT)和酶联免疫实验(ELISA)分别检测免疫后小鼠脾产生细胞因子IFN-γ的细胞数目和抗AFP抗体的水平。采用小鼠体内肿瘤负荷实验评价重组蛋白复合物的免疫效应。结果:经重组蛋白复合物免疫,ELISPOT和ELISA实验结果表明HSP70-AFP复合物免疫组分泌IFN-γ细胞因子的脾细胞数目和产生AFP的抗体水平显著高于AFP和HSP70免疫组(P<0.01)。HSP70-AFP复合物免疫组瘤体积也明显小于AFP和HSP70免疫组(P<0.01)。结论:研究证实了HSP70的免疫佐剂效应。实验结果表明重组HSP70-AFP复合物的连续免疫可以在小鼠体内产生有效的抗瘤免疫,HSP70-AFP重组蛋白复合物对于今后肿瘤疫苗的研究可能具有一定的意义。更多还原

【Abstract】 Objective:To construct a recombinant peptides complex-alpha-fetoprotein(AFP) and heat shock protein 70(HSP70),and study its ability to induce specific CTL response and its protective effect against AFP-producing tumor.Methods:A recombinant peptides complex was constructed by conjugating mouse alpha-fetoprotein(mAFP) with heat shock protein 70 protein.By way of intracutaneous injection,mice were primed and boosted with recombinant peptides complex HSP70-AFP,whereas single AFP or HSP70 injection as controls.ELISPOT and ELISA were used to test IFN-γ-producing splenocytes and the level of serum anti-AFP antibody from immunized mice respectively.In vivo tumor challenge was measured to assess the immune effect of the recombinant peptides complex.Results:By recombinant HSP70-AFP peptides complex immunization,the results of ELISPOT and ELISA showed that the number of splenic cells producing IFN-γ and the level of anti-AFP antibody of serum were significantly higher in HSP70-AFP group than those in AFP and HSP70 groups(P<0.01).The tumor volume in HSP70-AFP group was significantly smaller than that in AFP and HSP70 groups(P<0.01).Conclusion:The study confirmed the function of heat shock protein 70’s immune adjuvant.Sequential immunization with recombinant HSP70-AFP peptides complex could generate effective AFP-specific T cell responses and induce definite antitumor effects on AFP-producing tumors,which may be suitable for some clinical testing as a vaccine for HCC.更多还原