摘要
目的对胃癌细胞树突状细胞(dendtiticcells,DCs)融合疫苗激活杀伤性T淋巴细胞体内外肿瘤细胞杀伤活性进行研究,为胃癌DC疫苗免疫治疗提供实验依据。方法胃癌患者外周血单个核细胞经GMCSF,IL4,TNFα定向诱导获取DC。DC与SGC7901胃癌细胞经PEG诱导融合、HAT/HT筛选培养获得纯净融合细胞。MTT比色法检测融合疫苗体外杀伤胃癌细胞的细胞毒作用。观察融合疫苗对胃癌裸鼠种植瘤模型肿瘤生长的影响,流式细胞术AnnexinV/PI双标法检测瘤细胞凋亡,观察融合疫苗对肿瘤组织病理学改变的影响。结果①胃癌患者外周血单个核细胞体外定向诱导分化,可获得具备典型形态特征及表型的DC。②DC与SGC7901细胞经PEG诱导融合,HAT/HT筛选培养,可获得纯净融合细胞。③体外MTT比色检测,融合疫苗胃癌细胞杀伤率为(81.47±5.25)%,与混合培养DC组(70.53±2.46)%差异显著(P<0.05),与单纯T细胞组(21.67±2.55)%差异非常显著(P<0.01)。④融合疫苗组荷瘤裸鼠终末肿瘤体积平均(1.298±0.021)cm3,与对照组差异非常显著(P<0.01),抑瘤率为57.8%。⑤融合疫苗组肿瘤细胞凋亡率(54.9±1.38)%,坏死率为(21.04±1.47)%;与对照组瘤细胞凋亡百分率(20.02±0.39)%,坏死百分率(1.23±0.03)%比较差异非常显著(P<0.01)。⑥融合疫苗组肿瘤组织与对照组比较显著出血、坏死,瘤组织内大量淋巴细胞浸润。结论融合细胞疫苗激活T淋巴细胞体内外均可诱导显著胃癌细胞杀伤细胞毒活性;其应用于荷瘤动物体内,可以明显减慢肿瘤生长速度,诱导更多肿瘤细胞凋亡。
Objective To study on the cytotoxic effects in vivo and in vitro of the T lymphocytes which were activated by the DC-gastric cancer cell fusion vaccines. Methods The mononuclear cells were separated from the peripheral blood of gastric cancer patients and co-cultured with granulocyte-macrophage colony stimulating factors, interleukin-4 and tumor necrosis factor-α to generate mature dendritic cells. The human gastric cancer SGC7901 cells and the dendritic cells were fusioned by using polyethylene glycol, and the pure fusion cells were screened out by culturing with HAT and HT selective culture systems. The mixture of SGC7901 and DC without fusion and the T lymphocytes served as controls. The ability of fusion cells activated by T lymphocytes to kill SGC7901 gastric cancer cells was investigated by MTF in vitro. The anti-tumor effects were evalutated by detecting the growth of the new planted tumors in the nude mice, the apoptosis and proliferation of the planted cancer cells, the pathological changes of the tumor tissues and the prohibitive rate for planted tumors before and after the application of the fusion cell vaccine. Results The mature dendritic cells were obtained from peripheral blood mononuclear cells of gastric cancer patients successfully. Dendritic cells and SGC7901 cells were fusioned and the pure fusion cells were got. Fusion vaccine could induce strong anti-tumor biological effects in vivo and in vitro： Tumor growth was remarkably inhibited and the tumor size in mice receiving fusion cells was （ 1. 298 ± 0.021 ） cm^3, smaller than that in mice receiving T lymphocytes as treatment [ （ 2. 429 ± 0. 033） cm^2 ] （P 〈 0.01 ）. The inhibitory rate of the planted tumors was 57.8%. Detected by Annexin-V/PI double labeling flow-cytometry, the apoptosis rate of tumor cells in mice receiving fusion cells was （54.9 ± 1.38） %, higher than that in the mice receiving T lymphocytes [ （20.02 ± 0.39）% ] （P 〈 0.01 ）. The tumor tissues in the mice receiving fusion cells had more infiltrated lymphocytes, apoptotic cells, hemorrhage and necrosis than in the mice receiving T lymphocytes. Conclusion The fusion vaccine could inhibit the tumor growth and accelerate the apoptosis of tumor cells.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2006年第5期404-407,共4页
Journal of Third Military Medical University
关键词
胃癌
树突状细胞
融合疫苗
瘤细胞杀伤
gastric cancer
dendritic cell
fusion vaccine
cytotoxic effect
