论文部分内容阅读
目的:考察Hsp70L1对肿瘤细胞免疫原性的增强作用。方法:用RT-PCR的方法,从小鼠黑色素瘤B16细胞和C57BL/6小鼠脾脏中获得TRP2153-243及Hsp70L1基因。分别插入pcDNA3.1/V5-His真核表达载体,构建pHSP70L1、pTRP和pTRP2-Hsp3种表达载体。分别转染B16肿瘤细胞并制备坏死或凋亡瘤苗,免疫C57BL/6小鼠后移植B16肿瘤细胞,观察肿瘤生长曲线,采用流式细胞术(FCM)或微量细胞毒的方法检测荷瘤小鼠细胞因子INF-γ和CTL活性。结果:将经过HSP70L1、TRP2及TRP2-HSP基因修饰的坏死或凋亡的B16肿瘤细胞免疫正常小鼠后,观察到Hsp70L1及TRP2-Hsp基因修饰的坏死瘤苗可显著抑制荷瘤鼠肿瘤的生长,并显著促进荷瘤鼠脾脏淋巴细胞CTL活性和IFN-γ产生(P<0.05,P<0.01);HSP70L1免疫刺激作用在坏死瘤苗中更明显。结论:Hsp70L1可明显提高B16瘤苗的免疫原性,且对坏死细胞瘤苗的作用更显著。
Objective: To investigate the enhancement effect of Hsp70L1 on the immunogenicity of tumor cells. Methods: The TRP2153-243 and Hsp70L1 genes were obtained from mouse melanoma B16 cells and C57BL / 6 mouse spleen by RT-PCR. The recombinant plasmids were inserted into pcDNA3.1 / V5-His eukaryotic expression vector to construct expression vectors pHSP70L1, pTRP and pTRP2-Hsp. B16 tumor cells were transfected and necrotic or apoptotic tumor vaccine was prepared. B16 tumor cells were transplanted after C57BL / 6 mice were immunized. The tumor growth curve was observed. Flow cytometry (FCM) or trace cytotoxicity Murine cytokines INF-γ and CTL activity. Results: After necrosis or apoptosis of B16 tumor cells modified by HSP70L1, TRP2 and TRP2-HSP gene were induced in normal mice, it was observed that the necrotic tumor vaccine modified by Hsp70L1 and TRP2-Hsp gene could significantly inhibit tumor growth of tumor-bearing mice , And significantly promoted the CTL activity and IFN-γ production of spleen lymphocytes of tumor-bearing mice (P <0.05, P <0.01). The immunostimulatory effect of HSP70L1 was more obvious in necrotic tumor cells. Conclusion: Hsp70L1 can significantly enhance the immunogenicity of B16 tumor vaccine, and its effect on necrotic cell tumor vaccine is more significant.