【摘要】 目的:使用以毕赤酵母X-33为宿主菌,重组表达制备的rhHSP70与合成的HER2/neu抗原肽体外结合形成复合物,探讨其在肿瘤免疫治疗中的有效性。方法:在ADP存在的条件下,将rhHSP70与纯化的HER2/neu抗原肽体外非共价结合形成复合物,并分次免疫BALB/c小鼠,通过ELISPOT和颗粒酶释放法,检测该复合物诱导特异性CTL的能力。通过检测小鼠体内肿瘤的生长情况和肿瘤的组织切片,来观测该复合物对小鼠乳腺癌的免疫治疗作用。结果:rhHSP70-HER2/neu抗原肽复合物免疫小鼠后,可以增加T细胞分泌IFN-γ的能力,诱导出肿瘤特异性CTL,并对小鼠的乳腺癌有明显的治疗作用。结论:在毕赤酵母中重组表达的rhHSP70可以在体外与一定的抗原肽非共价结合形成复合物,该复合物具有较强的诱导特异性CTL的能力,可对实验鼠的乳腺癌起到明显的治疗作用。更多还原

【Abstract】 Objective: To investigate the possibility of using rhHSP70 obtained from the gene-modified Pichia pastoris and complexed with the CTL epitopes to cure cancer. Methods: We constructed rhHSP70-HER2/neu peptides complexes in vitro and immunized BALB/c mice with breast cancer, then assayed the specific CTL induced by the complexes through ELISPOT and Granzyme releaseing method. At the same time, we observed the tumor volume and the histology of the tumors to analyze the remedial effect of the complexes. Results: The rhHSP70-HER2/neu peptide complexes could enhance T cells to secret IFN-γ, induce specific CTL and had obvious remedial effect on mice with breast cancer. Conclusion: rhHSP70 produced by Pichia pastoris can bind some polypeptides in vitro and the subsequent rhHSP70-peptide complexes have therapeutic effects on mice with breast cancer.更多还原