热休克胃癌细胞负载DC-CIK细胞杀伤癌细胞活性的研究

目的研究负载相关抗原的树突状细胞(DC)联合细胞因子诱导的杀伤(CIK)细胞对胃癌细胞的杀伤作用。方法取健康人外周血并分离出外周血单个核细胞(PBMC),经不同细胞因子分别诱导出DC和CIK细胞,热休克方法处理胃癌MKN-28细胞株并制备肿瘤抗原用于负载DC,与CIK细胞共培养,以流式细胞术检测DC、CIK细胞表型,MTS法检测CIK细胞对MKN-28细胞的杀伤作用。结果 CIK与DC共培养后,与单纯CIK细胞相比较,增殖活性明显提高,CD3+CD8+、CD3+CD56+双阳性细胞数增多并且具有更强的杀瘤活性,效靶比为20.0∶1时,负载MKN-28抗原的DC-CIK(Ag-DC-CIK)组对MKN-28的杀伤活性为(57.96±2.23)%,远大于未负载MKN-28抗原的DC-CIK组[(38.02±2.06)%]和单纯CIK组[(29.78±1.84)%],差异具有统计学意义(P<0.05)。结论以热休克凋亡肿瘤细胞抗原负载的DC能促进CIK细胞增殖分化,并提高其对胃癌细胞的杀伤活性。

Study on the killing activity of DC-CIK cells pulsed with gastric cancer cells of heat shock against cancer cells

Objective To study the killing effect of dendritic cells（DC） and cytokine-induced killer（CIK） cells pulsed with specific antigens against gastric cancer cells.Methods Peripheral blood mononuclear cells（PBMC）were isolated from healthy subjects.DC and CIK cells were induced by different cytokines.Gastric cell line MKN-28 lysates of heat shock were used as antigen and pulsed DC,which co-cultured with CIK cells.The phenotypes of DC and CIK cell membranes were determined by flow cytometry,and the killing activity of CIK against MKN-28 was determined by（MTS）.Results Compared with CIK,the co-cultured DC-CIK presented a significantly higher proliferation.The numbers of CD3＋CD8＋ and CD3＋CD56＋ cells increased,and the killing effect was enhanced.The cytotoxic activity to MKN-28 of DC-CIK pulsed with MKN-28 antigen（Ag-DC-CIK） group was（57.96±2.23）%,which was more strong than（38.02±2.06）% of DC-CIK pulsed without MKN-28 antigen group and（29.78±1.84）% of CIK group,respectively [the ratio of effect to target =20.0∶ 1].Conclusions Heat shock tumor-lysate-pulsed DC can strengthen the proliferation and killing activity of CIK against gastric cancer cells.