热休克肿瘤细胞抗原负载的树突状细胞瘤苗对原位接种结肠肿瘤的免疫预防作用  

Prophylactic effects of dendritic cells pulsed with lysates of heat-shocked tumor cells in mice with colonic inoculation of tumor cells

作　　者：邱健[1] 李国威[1] 隋延仿[2] 宋宏萍[2] 司少艳[2] 葛伟[2] 

机构地区：[1]西安交通大学第二医院普外科,陕西省西安市710004 [2]中国人民解放军第四军医大学病理学教研室,陕西省西安市710032

出　　处：《世界华人消化杂志》2005年第5期635-639,共5页World Chinese Journal of Digestology

基　　金：陕西省自然科学基金资助项目;No.2004C271~~

摘　　要：目的:研究热休克肿瘤细胞抗原负载的树突状细胞的体内抗肿瘤作用以及对原位接种结肠肿瘤小鼠的免疫保护作用. 方法:肿瘤细胞CT-26经热休克处理后制备冻融抗原,体外负载树突状细胞,免疫同源小鼠(热休克组).以IFN-γELISPOT实验、LDH释放实验检测小鼠体内肿瘤特异性T细胞免疫反应,以原位接种结肠肿瘤的体积以及荷瘤小鼠的生存时间来评价该树突状细胞瘤苗对结肠原位接种CT-26小鼠的免疫保护作用,并与非热休克肿瘤细胞抗原负载的树突状细胞的免疫效果(非热休克组)作一比较. 结果:热休克处理肿瘤细胞可提高肿瘤细胞抗原负载的树突状细胞表面MHC-Ⅱ类分子(66.3% vs 59.1%)、共刺激分子CD86的表达(39.4% vs 36.7%);热休克组小鼠脾淋巴细胞中肿瘤特异性CTL数量高于非热休克组(P=0.001),相同效靶比下前者肿瘤特异性细胞毒活性强于后者:热体克组与非热休克组小鼠原位接种结肠肿瘤14 d后的肿瘤体积均明显小于单纯DC免疫组(P=0,000),但HSCT-26DC免疫组与CT-26 DC免疫组间差异无显著性(P=0.480).单纯DC免疫组50% (3/6)可见腹膜转移,CT-26 DC免疫组以及HSCT- 26DC免疫组均未见腹腔转移;热休克组小鼠荷瘤生存时间显著长于非热休克组(P=0.0384). 结论:热休克处理肿瘤细胞可以提高肿瘤细胞抗原负载的树突状细胞的体内抗肿瘤作用,热休克肿瘤细胞抗原负载的树突状细胞瘤苗对原位接种结肠肿瘤小鼠具有良好的免疫保护作用.AIM: To investigate the antitumor effect of dendritic cells (DC) pulsed with lysates of heat-shocked tumor cells and the prophylactic effects of this DC tumor vaccine in mice inoculated with tumor cells in colon. METHODS: CT-26 cells were heat-shocked at 42 ℃ for 1 h (HSCT-26) and then frozen-thawed repeatedly to lyse. Bone marrow-derived DCs pulsed with the lysate were used to immunize BALB/c mice. Using IFN-γ enzyme-linked immunospot (ELISPOT) and LDH release assay, the anti-tumor response of cytotoxic T lymphocytes (CTLs) was evaluated. The immunoprophylactic effect induced by heat-shocked CT-26 cell lysate pulsed DC (HSCT-26 DC) in colon cancer mouse model was compared with that induced by non-heat-shocked CT-26 cells lysate pulsed DC (CT-26 DC) on tumor volume and survival time. RESULTS: Compared with CT-26 lysates, pulsing with HSCT-26 lysate enhanced the expression of MHC-II molecule (66.3% vs 59.1 %) and co-stimulating molecule CD86 (39.4% vs 36.7%) on DC surface. The quantity of CTLs induced by HSCT-26 DC was more than that induced by CT-26 DC (P= 0.001). The specific cytotoxic activity of CTLs induced by HSCT-26 DC was more potent than that of CTLs induced by CT-26 DC at the same E : T ratio. Fourteen days after colon inoculation of CT-26 cells, the tumor volume of HSCT-26 DC immunized mice was similar to that of CT-26 DC immunized mice (P= 0.480), and both were much smaller than that of DC immunized mice (P = 0.000). 50% of DC immunized mice suffered from peritoneal metastasis, whereas none of HSCT-26 DC immunized mice and CT-26 DC immunized mice was involved. The survival time of colon cancer model mice immunized with HSCT-26 DC was significantly longer than that of the mice immunized with CT-26 DC (P= 0.0384). CONCLUSION: Pre-heated tumor cells can enhance the antitumor effects elicited by tumor cell lysate pulsed DCs in vivo. Vaccination with DCs pulsed with lysates of heat-shocked tumor cells can prolong the survival time of mice with colon cancer by effectively suppressing the growth of colon c

关 键 词：树突状细胞瘤苗 肿瘤细胞 原位接种 抗原负载 结肠肿瘤 免疫预防作用 体内抗肿瘤作用 免疫保护作用 MHC-Ⅱ类分子 肿瘤特异性CTL 热休克处理 ELISPOT 细胞免疫反应 肿瘤小鼠 生存时间 IFN-γ 共刺激分子 脾淋巴细胞 细胞毒活性 

分 类 号：R735.35[医药卫生—肿瘤]