树突细胞-食管癌融合细胞体外诱导特异性抗食管癌免疫反应

背景与目的:树突细胞(dendritic cells,DC)是人体专职的抗原呈递细胞,以DC为基础的DC/肿瘤细胞融合疫苗可以有效地诱导特异性抗肿瘤免疫应答.本研究旨在探讨成熟的DC与人食管癌细胞株ECl09细胞融合疫苗体外诱导特异性抗食管癌细胞的免疫反应.方法:从健康志愿者外周血中分离出单个核细胞(peripheral blood mononuclear cells,PBMC),在重组人粒/巨噬细胞集落刺激因子(recombinant human granulocyte-macrophage-colony-stimulating factor,rhGM-CSF)和白介素(interleukin-4,IL-4)作用下体外诱导DC,采用聚乙二醇(polyethylene glyco1,PEG)融合法使DC与EC109细胞融合制备DC/EC109细胞融合疫苗,四甲基偶氮唑盐(MTT)实验检测融合疫苗刺激T淋巴细胞增殖的活性,乳酸脱氢酶(LDH)实验检测融合疫苗活化的细胞毒性T淋巴细胞(cytotoxic T lymphocyte,CTL)在体外特异性杀伤EC109细胞的活性,并与对人胃癌细胞株SGC7901及人乳腺癌细胞株MCF7的杀伤作用进行比较.结果:DC与EC109细胞的融合效率最高达到22.25%.DC/EC109细胞融合疫苗能有效地刺激T淋巴细胞的增殖反应,其刺激T淋巴细胞增殖的能力显著高于DC或EC109细胞(P＜0.05).DC/EC109细胞融合疫苗活化的CTL对EC109细胞具有特异性的杀伤作用,对EC109细胞的杀伤作用显著强于对SGC7901细胞及MCF7细胞的杀伤作用(P＜0.05).结论:DC/EC109细胞融合疫苗能有效诱导抗EC109细胞的特异性免疫应答.

Antitumor efficacy of fusion cells from esophageal carcinoma cells and dendritic cells as a vaccine in vitro

BACKGROUND & OBJECTIVE: Dendritic cells (DCs) are antigen-presenting cells, and DC-based fusion vaccine of DCs with tumor cells can induce specific immune response against tumor cells effectively. This study was to investigate the antitumor immunity efficacy of fusion vaccine of DCs with human esophageal carcinoma EC109 cells in vitro. METHODS: Peripheral blood mononuclear cells (PBMCs) from healthy volunteers were isolated, and cultured with recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and interleukin-4 (IL-4) to generate DCs. Fusion cells of DCs with EC109 cells were generated by polyethylene glycol (PEG) protocol. The T-cell proliferation response stimulated by DC/EC109 cells was detected by MTT assay. The killing efficacy of cytotoxic T lymphocytes (CTLs), activated by DC/EC109 cells, on EC109 cells was evaluated by LDH assay in vitro, and compared with the killing efficacy on human gastric carcinoma SGC7901 cells and human breast cancer MCF7 cells. RESULTS: The highest fusion efficiency of DCs with EC109 cells was 22.25%. The stimulating efficacy of DC/EC109 cells on the proliferation of T cells was significantly higher than those of DCs and EC109 cells (P<0.05). DC/EC109 cells induced specific CTLs against EC109 cells, and the killing efficacy of the CTLs was significantly higher for EC109 cells than for SGC7901 cells or MCF7 cells (P<0.05). CONCLUSION: DC/EC109 fusion vaccine can induce specific antitumor response against EC109 cells effectively.