体外大量制备新型gp96肿瘤疫苗及其诱导的特异性抗肿瘤免疫应答分析

Large-scale in vitro preparation of new gp96 tumor vaccine and analysis of its induction of specific anti-tumor immunoresponses

旨在体外组装酵母菌表达的gp96(Recombinant gp96,rgp96)蛋白与B16.F10黑色素瘤抗原,大量制备新型gp96肿瘤疫苗,并研究其诱导的特异性抗肿瘤免疫应答。利用体外组装的rgp96-肿瘤抗原复合物免疫C57BL/6小鼠,并通过酶联免疫斑点实验、细胞因子染色、杀伤实验技术进行分析,结果显示与单纯rgp96或肿瘤抗原免疫组相比,体外组装的rgp96-肿瘤抗原复合物免疫能够显著抑制B16肿瘤的生长,而且能够明显提高肿瘤特异性T细胞活性。rgp96-肿瘤抗原复合物的抗肿瘤免疫活性与从肿瘤组织中提取的gp96接近。研究结果为大量制备新型gp96肿瘤疫苗提供了依据。

Heat shock protein gp96 isolated from tumor tissues holds great promise for tumor immunotherapy.However,at present only very limited amount of gp96 protein can be isolated from tumor tissues.Here,we reconstituted the yeast-expressed gp96（recombinant gp96,rgp96） with B16.F10 melanoma antigens in vitro to prepare new gp96 tumor vaccine on large-scale,and analyzed its induction of specific anti-tumor immunoresponses by ELISPOT,IFN-γ intracellular staining and cytotoxicity assays.Immunization with rgp96-tumor antigen complexes significantly inhibited B16 tumor growth compared with either rgp96 or tumor antigens alone and led to enhancement of tumor-specific T-cell activities,which was found similar to that of tumor tissue derived gp96.Our results therefore may provide bases for large-scale preparation of the new generation of gp96 tumor vaccines.