摘要
目的探讨溶血性链球菌制剂(OK432)联合肿瘤冻融抗原诱导小鼠骨髓树突状细胞(DCs)成熟的作用,改进体外诱导DCs成熟的方案,为后期制备临床使用的抗肿瘤DCs疫苗提供新的技术路线。方法分离小鼠骨髓单核细胞(BMMCs),并在含10%胎牛血清(FCS)、重组小鼠粒-巨噬细胞集落刺激因子(rmGM-CSF)、重组小鼠白细胞介素-4(rmIL-4)的培养体系中诱导培养,部分加入OK432和/或肿瘤冻融抗原冲击,光镜和透射电镜下观察细胞形态,流式细胞仪检测细胞表面分子CD83和CD86的表达情况。结果经OK432联合肿瘤细胞冻融抗原冲击的DCs形态学特征典型。肿瘤冻融抗原冲击后,DCs表面CD83和CD86的表达上调,OK432进一步刺激后,CD83和CD86的表达率显著升高。结论采用OK432联合肿瘤冻融抗原诱导DCs成熟的方案相对简便;OK432能有效促进肿瘤冻融抗原冲击后的骨髓DCs进一步成熟。
Objective To investigate the effect of OK432 and freeze-thawed tumor cell antigen in maturation of dendritic cells（DCs） from mouse bone marrow, and improve the means of maturation of den- dritic cells in vitro so as to offer a new technical way of preparation DCs-based vaccines for the cancer im- munotherapy. Methods Mouse bone marrow mononuclear cells （BMMCs） were cultured in RPMI1640 containing 10% FCS,rmGM-CSF and rmlL-4, pulsed with OK432 and /or freeze-thawed tumor cell anti- gen. Morphological characters of DCs were observed by the optical microscope and the electron micro- scope; DCs surface molecules CD83 and CD86 were assayed by flow cytometry. Results Induced by OK432 and freeze-thawed tumor cell antigen, DCs showed morphologically typical characters. And they expressed much more antigens CD83 and CD86 on the surface after cultured0by pulsed with freeze-thawed tumor cell antigen. Notably more antigens CD83 and CD86 were detected on the surface of DCs which were pulsed with both of OK432 anf freeze-thawed tumor cell antigen. Oonclusion It was simple and practicable to induce maturation of DCs by OK432 and freeze-thawed tumor cell antigen. OK432 could markedly enhanced the maturity of DCs which had been pulsed with freeze-thawed tumor cell antigen.
出处
《福建医科大学学报》
2008年第5期396-399,407,共5页
Journal of Fujian Medical University
基金
福建省自然科学基金资助项目(C0410020)
