论文部分内容阅读
目的探讨热休克瘤细胞裂解物冲击的异基因树突状细胞(DC)抗肿瘤免疫效应。方法应用热休克(43℃,1h)小鼠 Lewis 肺癌细胞(LLC)裂解物(LhTCL)冲击的小鼠骨髓细胞衍生的树突状细胞(DC)作为瘤苗(DC/LhTCL)。应用 Western 印迹检测可诱导 HSP70表达。用 FITC-标记的 LhTCL 和 CFSE-标记的异基因 DC 分别进行体外吞噬和体内示踪实验,藉酶联免疫吸附(ELISA)、流式细胞术(FCM)和共聚焦荧光显微镜观察分析不成熟异基因 DC 体外摄取 LhTCL 对其表型和 IL-12产生的影响以及受鼠接种局部引流淋巴结DC(LD-DC)对异基因 DC 的摄取。然后,通过免疫接种的小鼠脾细胞体外诱生 CTL 的杀瘤活性(LDH法)和产生的 Th-1相关细胞因子检测,LLC移植瘤的免疫预防和早期带瘤鼠的治疗实验以评价 C57BL小鼠接种异基因或同系 DC/LhTCL 瘤苗的产生的抗肿瘤免疫效应。结果 (1)热休克增加 LLC 胞浆内可诱导 HSP70表达。(2)小鼠不成熟异基因 DC 能充分摄取 LhTCL 并促进其 DC 成熟(增加 DC 表面共刺激分子表达和 IL-12分泌)。(3)皮下注射 LhTCL 冲击的异基因 DC 观察到有促进受鼠 DC 摄取异基因 DC 的效应。(4)LhTCL 冲击的异基因 DC 免疫的 C57BL 小鼠脾细胞能特异性杀伤活 LLC,分泌 Th-1相关的细胞因子(高水平IFN-γ和低水平 IL-4和 IL-10)。(5)C57BL 小鼠 LLC 移植瘤免疫保护实验和治疗实验均证实,免疫接种异基因 DC/LhTCL 瘤苗能够阻止或延缓 LLC 肿瘤的生长,其产生的免疫保护效应要比同系 DC/LhTCL 瘤苗强。结论接种热休克处理的肿瘤裂解物冲击的异基因树突状细胞瘤苗有促进抗肿瘤免疫的效应。
Objective To investigate the anti-tumor immune effect of allogeneic dendritic cells (DCs) impacted by heat shock tumor cell lysate. Methods Mouse bone marrow cell-derived dendritic cells (DCs) impacted by heat shock (43 ° C, 1 h) mouse Lewis lung carcinoma (LLC) lysate (LhTCL) were used as tumor vaccine (DC / LhTCL). Western blot detection can induce HSP70 expression. In vitro phagocytosis and in vivo tracing experiments were performed with FITC-labeled LhTCL and CFSE-labeled allogeneic DCs respectively. Immature isoforms were analyzed by enzyme-linked immunosorbent assay (ELISA), flow cytometry (FCM) and confocal fluorescence microscopy Effect of in vitro uptake of LhTCL by gene DC on its phenotype and IL-12 production and uptake of allogeneic DC by murine inoculation of locally draining lymph node DC (LD-DC). Then, the cytotoxic activity of CTL (LDH method) and the detection of Th-1-related cytokines produced by the immunized mouse spleen cells were induced in vitro, the immunoprecipitation of LLC xenografts and the treatment experiment of early-stage tumor-bearing mice to evaluate C57BL Anti-tumor immune effects of mice vaccinated with allogeneic or syngeneic DC / LhTCL vaccines. Results (1) Heat shock increased LLC cytoplasmic inducible HSP70 expression. (2) Mouse immature allogeneic DC can fully ingest LhTCL and promote its DC maturation (increase DC surface costimulatory molecule expression and IL-12 secretion). (3) The effect of subcutaneous injection of allogeneic DCs with LhTCL impact on allogeneic DC induced by DC in mice was observed. (4) The spleen cells of C57BL mice immunized with LhTCL and allogeneic DCs could specifically kill live LLC and secrete Th-1 related cytokines (high level of IFN-γ and low levels of IL-4 and IL-10). (5) C57BL mouse LLC xenograft tumor immunization experiments and treatment experiments all confirmed that immunization of allogeneic DC / LhTCL vaccine can prevent or delay the growth of LLC tumor, and its immune protective effect than the homologous DC / LhTCL vaccine Strong. Conclusion Allogeneic dendritic cell vaccine impacted by heat shock treated tumor lysate can promote anti-tumor immunity.