目的:探讨负载肿瘤抗原的树突状细胞(DC)疫苗诱导的细胞毒性T淋巴细胞(CTL)的体外杀瘤作用及对荷瘤小鼠的治疗作用.方法:体外培养的小鼠骨髓树突状细胞用小鼠结肠腺癌细胞株CT26细胞抗原冲击致敏制备负载肿瘤抗原的DC疫苗;负载肿瘤抗原的DC疫苗激活同源T淋巴细胞产生细胞毒性T淋巴细胞(CTL),采用乳酸脱氢酶(LDH) 4 h释放法检测CTL在体外对CT26细胞的杀伤活性;建立CT26荷瘤小鼠模型,应用CTL治疗荷瘤小鼠,观察肿瘤大小和小鼠存活期.结果:负载CT26肿瘤抗原的DC疫苗能够诱导T细胞增殖分化为肿瘤特异CTL,该CTL对CT26细胞有高效而强烈的杀伤作用,杀伤率为(83.95±11.25)%,而对B16细胞、3LL Lewis 细胞无明显杀伤作用,杀伤率分别为(12.75±5.36)%和(11.38±4.57)%.应用CTL治疗荷瘤小鼠能显著抑制荷瘤小鼠肿瘤的生长,延长荷瘤小鼠存活期.结论:负载肿瘤抗原的DC疫苗能够诱导高效而特异的CTL杀瘤活性并能治疗荷瘤小鼠.提示负载肿瘤抗原的DC疫苗诱导的CTL可能在肿瘤的免疫治疗中发挥重要作用.
Objective: To explore the anti-tumor effects of cytotoxic of T-lymphocytes (CTL) induced by dendritic cells (DC) vaccine loaded with tumor antigen. Methods: Murine bone marrow DC were induced and pulsed with murine CT26 colorectal adenocarcinoma cells antigen to prepare DC vaccine loaded with tumor antigen. The tumor specific CTL was generated form activated autologous T cell by DC vaccine loaded with tumor antigen, and the killing activity of CTL to CT26 cells was tested by the lactate dehydrogenase (LDH) release assay in vitro. CT26 colorectal adenocarcinoma-bearing mice model were established and treated with the tumor specific CTL, the tumor size and the survival period of mice were monitored. Resuit: CTL induced by DC vaccine loaded with tumor antigen could selectively kill CT26 cells [killing rate: (83.95+11.25) %], but slightly kill B16 cells and 3LL Lewis cells [Killing rate: (12.75+5.36)%, (11.38 +4.57)%, respectively]. The tumor specific CTL could significantly inhibit the growth of implanted tumor in mouse and prolong the survival period of tumor-bearing mice. Conclusion: The DC vaccine loaded with tumor antigen could induce efficient and specific killing-tumor activity of CTL, and the tumor specific CTL could treat tumor-bearing mice. The results suggest that the specific CTL induced by DC vaccine loaded with tumor antigen may play an important role in tumor immonotherapy.