摘要
目的探讨体外扩增树突状细胞(DC)的方法,体外研究DC介导的细胞毒性T淋巴细胞(CTL)对小鼠胰腺癌的特异性杀伤作用。方法联合应用重组鼠源性粒细胞-巨噬细胞集落刺激因子(GM-CSF)和IL-4体外诱导骨髓源性DC,与肿瘤细胞溶解物共培养制备DC疫苗;观察DC在负载抗原后体外诱导的主动特异性CTL对胰腺癌细胞的杀伤作用。结果用GM-CSF和IL-4成功地在体外扩增了骨髓源性DC。TNF-α可诱导骨髓源性DC成熟,使其表达的CD54、主要组织相溶性复合物-Ⅱ、CD86等表面分子明显升高,并增强自分泌IL-12和抗原的递呈能力,与对照组比较,差异有统计学意义(P<0.05)。未负载肿瘤细胞抗原的DC所冲击的T淋巴细胞不能有效地识别特定的靶细胞并产生杀伤作用,DC通过捕获并递呈坏死肿瘤细胞抗原才能诱发显著的主动特异性CTL,体外对胰腺癌细胞的杀伤效果非常显著(P<0.01)。结论TNF-α可诱导DC的成熟,使DC的抗原递呈和自分泌IL-12的能力显著提高,DC递呈抗原后能诱发显著的主动特异性CTL。
Objective To study the method of obtaining a large number of dendritic cells （DC）. To study the specific cytotoxicity T lymphocyte （CTL） effect against tumor cells initiated by DC pulsed with peptide of cancer cell. Methods Development of cells with cytologic features of DC in bone marrow cultures supplemented with granulocyte macrophage-colony stimulus factor （GM-CSF） and IL-4. Determining the DC phenotype and the specific structure by electronic microscopy. The CTL effect against pancreatic carcinoma leading by the DC pulsed with tumor cells lysate in vitro was observed. Results A large number of typical DC was proliferated by supplementing with GM-CSF and IL-4 cytokines. DC had specific cell appearance and structure, and highly expressed various cell surface molecules. TNF-α had the ability of stimulating DC mature, the mature DC had the enhancing abilities of antigen presenting and IL-12 self-secreting, as well as, expressed higher levels of CD54, MHC- Ⅱ and CD86 molecules than control group （P〈0.05）. T lymphoid cell stimulated by DC without tumor antigen could not recognize and kill the target cells, only if DC pulsed with peptide of cancer cell can lead a strong immune response to special tumor cells. The inhibiting ratio of CTL was significantly higher than that in other groups （P〈0.01）. Conclusion Bone marrow DC has strong ability of inducing special CTL against determined cancer cells after they are pulsed with tumor cell lysate. DC vaccine is probably a new immunotherapeutic method against tumor in the near future.
出处
《中国普外基础与临床杂志》
CAS
2006年第6期689-692,共4页
Chinese Journal of Bases and Clinics In General Surgery
基金
辽宁省科技厅自然基金资助项目(编号:20042138)~~
关键词
树突状细胞
胰腺癌
免疫反应
Dendritic cell Pancreatic carcinoma Immune response
