沙培林诱导脐血树突状细胞的成熟

Sapylin in Maturation of Dendritic Cells from Umbilical Cord Blood

目的改进体外诱导脐血树突状细胞(DC)成熟的方案,为后期制备临床使用的抗肿瘤DC疫苗提供新的技术路线。方法分离脐血单个核细胞(CBMC)并在含10%同源脐血浆、重组人粒-巨噬细胞集落刺激因子(rhGM-CSF)和重组人白细胞介素-4(rhIL-4)的培养体系中诱导培养,部分加入肿瘤冻融抗原和/或沙培林(OK-432)冲击,光镜和电镜下观察细胞形态,流式细胞仪检测培养前后各组细胞表面抗原CD1a和CD83的表达情况。结果培养过程中,细胞形态发生明显变化,呈现典型的DC形态。收集第9天各组DC,细胞表面均高度表达CD1a和CD83,与贴壁分离的CBMC比较有统计学意义,肿瘤冻融抗原冲击后,DC表面CD1a和CD83的表达上调,沙培林进一步刺激后,CD1a和CD83表达率显著升高。结论诱导脐带血来源的前体细胞分化为未成熟DC的途径相对简便;沙培林能促进肿瘤冻融抗原冲击后的脐血DC进一步成熟。

Objective To investigate the means of maturation of dendritic cells（DC） from umbilical cord blood in vitro so as to offer a new technical way of preparation DC-based vaccines for the cancer immunotherapy. Methods Human cord blood mononuclear cells（CBMC） were cultured in RPMI 1640 containing 10% autologous plasma, GM-CSF and IL-4, some of which were supplemented with tumor lysates and/or sapylin（OK-432）, and then observed by the optical microscope and the electron microscope. Flow cytometry was used to detect DC surface antigens before and after culture. Results Cells changed markedly in culture and appeared typical morphology of DC. And they expressed much more antigens CDla and CD83 on the surface after culture by pulsed with tumor lysates. More antigens were detected on the surface of DC when treated with both of tumor lysates combined with sapylin. Conclusion Immature DC can be induced from human CBMC. Sapylin could markedly enhanced the maturity of DC which had been treated with tumor lysates.