【作者】 范立侨； 李勇； 赵增仁； 杨进强； 赵群； 刘羽； 王力利； 宋振川； 张志栋；

【Author】 Zhao Zeng-Ren LI Yong Yang Jin-Qiang et al the Third Department of Surgery, the Fourth Hospital of Hebei Medical University, Shijiazhuang, 050011

【机构】 河北医科大学第四医院暨河北省肿瘤医院外科；

【摘要】 目的:探讨胃癌细胞抗原致敏的DC诱导T淋巴细胞增殖和杀伤胃癌细胞的效率,为胃癌的免疫生物治疗提供新的治疗途径和实验依据。方法:自健康人外周血诱导培养获取成熟DC,自同一人提取外周血、扩增T淋巴细胞,流式细胞术检测其CD3、CD4、CD8及CD56表型。以人胃癌 OCUM-2MD3细胞株冻融抗原致敏DC,MTT法检测致敏 DE对T细胞增殖的影响和致敏DC活化的特异性CTL对胃癌细胞的体外杀伤效率。结果1．人外周血中提取的PBMC, 在体外培养,经rhIL-2、rhGM-CSF刺激后,可获大量表达CD3的T淋巴细胞。2．经MTT法检测胃癌细胞抗原致敏 DC对人外周血T淋巴细胞增殖的影响,可见致敏DC组的 SI值为0．573±0．115,未致敏DC组为0．183±0．074、对照组为0．185±0．026、空白组为0．176±0．053,致敏DC 组刺激指数显著高于其它三组,差异有显著性(P<0．01)。 3．经FCM检测,胃癌细胞抗原致敏DC主要刺激人外周血 T淋巴细胞增殖为CD8+表达阳性的CTL(60．58±8．43) ％,显著高于其它组,差异有显著性(P<0．01)。4．与未致敏DC组和对照组相比,经胃癌细胞抗原致敏DC激活的T 淋巴细胞对胃癌细胞的杀伤效应显著增加,差异有显著性(P <0．01)。并且随效靶比增加,杀伤效应亦显著增加(P<0． 01)。结论:自外周血中提取的PBMC,在体外培养扩增可获大量T淋巴细胞。胃癌细胞抗原致敏DC可显著刺激T淋巴细胞增殖为CD8+细胞毒CTL。胃癌细胞抗原致敏DC活化的特异CTL在体外具有高效杀伤胃癌细胞的作用。本研究为胃癌的免疫生物治疗提供了新的治疗途径和实验依据。更多还原

【Abstract】 Objective: To investigate the induction of DCs sensitized by gastric cell antigen to amplification and killing efficiency of T lymphocytes, and provide experiment foundation and new therapeutic methods for immunobiologic treatment of gastric cancer. Methods: Mature DCs were obtained through induction and culture from peripherial blood of healthy human, T lymphocytes were extracted and amplified from peripherial blood sample collected from the same man, and their phenotypes CD3, CD4, CD8 and CD56 were assayed by flow cytometry. After allergized with freeze thawing antigen of human gastric cancer cell line OCUM-2MD3, the effect of DCs on the amplification of T cells was assayed by MTT. Meanwhile the killing efficency of specific CTL, which activated by sensitized DCs, to gastric cancer cells in vitro was detected by MTT. Results: 1 Lots of T lymphocytes expressing CD3 could be got from PBMCs extracted from human peripheral blood through culture in vitro and stimulation of rhIL-2 and rhGM-CSF. 2 MTT assay showd that DCs sensitized by gastric cancer antigen could affect the proliferation of T lymphocytes. SI was 0.573 ± 0.115 in sensitized DCs group, 0.183 ± 0.074 in non-sensitized DCs group, 0.185 ± 0.026 in control group and 0.176 ± 0.053 in blank group. SI of sensitized DCs group was higher evidently than it of other groups, the difference was statistically significant (p<0.01). 3 FCM assay showed that DCs sensitized by gastric cancer antigen mainly stimulated T lymphocytes in human peripheral blood to proliferate into CD8+ CTL (60.58 ± 8.43)%, it was higher than other groups, the difference was statistically significant (p<0.01). 4 The killing efficency of T lymphocytes to gastric cancer cells increased obviously after activated by DCs sensitized by gastric cancer cell antigen. Compared with it of non-sensitized DCs group and control group, the difference was statistically significant (p<0.01). and moreover the killing efficency of T lymphocytes could increase following the rise of ratio of effect and target cells (p<0.01). Conclusion: Lots of T lymphocytes could be got from PBMCs extracted from human peripheral blood through culture in vitro. DCs sensitized by gastric cancer antigen could stimulate T lymphocytes to proliferate into CD8+ CTL The specific CTLs activated by DCs sensitized with gastric cancer antigen could effectively kill gastric cancer cells in vitro. These results provided experimental foundation and new therapeutic methods for immunobiologic treatment of gastric cancer.更多还原