OK-432增强脐血树突状细胞瘤苗抗胃癌作用的实验研究

OK-432 enhances the effect of umbilical cord blood dendritic-cells treated with gastric cancer tumor vaccine

目的:用脐血浆代替胎牛血清,观察OK-432对脐血来源的树突状细胞(DCs)功能的影响,探讨制备高效抗肿瘤DCs疫苗的优化方案。方法:分离脐血单个核细胞并在含10%同源脐血浆、GM-CSF和IL-4的培养体系中诱导培养,部分加入肿瘤冻融抗原和/或OK-432冲击,MTT法检测DCs体外刺激同种混合淋巴细胞增殖的免疫活性,LDH法检测DCs诱导的CTL对不同靶细胞的细胞毒作用。结果:收集第9 d的各组DCs,肿瘤冻融抗原冲击后的DCs呈成熟形态,能呈递胃癌相关抗原给淋巴细胞刺激其增殖并诱导对胃癌细胞株特异的CTL毒性。OK-432联合肿瘤冻融抗原冲击能增强其抗原呈递功能,诱导更强的混合淋巴细胞增殖反应及针对胃癌细胞株的强烈CTL杀伤效应。结论:该DCs培养方案具有培养时间短、细胞因子消耗少、试剂简单等优点,可用于制备DCs肿瘤疫苗,有临床应用价值。

AIM： To evaluate the effect of replacing fetal cattle serum with umbilical cord plasma and application of OK -432 on dendritic cells from umbilical cord blood in vitro so as to offer a new technical way of preparation high - powered dendritic - cell - based vaccines for the cancer immunotherapy. METHODS ： Human cord blood mononuclear cells （CBMCs） were cultured in RPMI- 1640 containing 10% autologous plasma, GM -CSF and IL-4, some of which were supplemented with tumor lysates and/or OK -432. MTT assay was applied to measure the antigen presenting ability of DCs in allo - MLR. Killing rates of autologous T lymphocytes induced by DCs on different target cells were measured by LDH method. RESULTS ： Cells appeared typical morphology of DCs after culture and the allo - stimulate capacity of DCs and the CTL response in vitro were enhanced by treating with tumor lysates and OK -432. CONCLUSION： Mature DCs can be induced from human CBMCs by this means with fewer cytokines and less time. The tumor lysate antigens are captured by DCs treated with tumor lysates and OK -432, and presented to lymphocytes successfully, indicating a new way to develop dendritic - cell - based vaccines for clinical immunotherapy of gastric cancer and other tumors.