摘要
目的 以实体瘤患者外周血造血干细胞为来源 ,探讨体外诱导更成熟、功能更强的树突状细胞 (DC)的方法。方法 以血细胞分离机分离经动员的外周血造血干细胞 (PBSC) ,加入rhGM CSF、rhIL 4和rhTNF α培养 9d后分两组 ,其中一组洗去含细胞因子的培养液 ,并给予钙离子载体 (CI)A2 3187处理 2 4h ,而另一组仍维持原培养液培养 2 4h ;对所获得的两组细胞分别进行细胞形态学、细胞表面抗原及刺激同种异体混合淋巴细胞反应 (allo MLR)能力的分析。结果 两组细胞在倒置显微镜和扫描电镜下均显示典型的树突状细胞形态 ,流式细胞仪分析均高表达CD1α、HLA DR、CD80和CD4 0分子 ;但给予A2 3187进一步处理 2 4h的细胞比无A2 3187处理的细胞表达更丰富的CD86和CD83分子及具有更强的刺激allo MLR的能力。结论 组合细胞因子培养获得的PBSC来源的DC ,经CI进一步诱导后可获得更成熟及功能更强的DC。
Objective To explore a culture method for generating functional and mature dendritic cells (DCs) from human peripheral hematopoietic stem cells of patients with solid tumor. Methods After stem cell mobilization, the peripheral blood stem cell (PBSC) was isola- ted from the blood sample by blood corpuscle separator. Then the PBSCs were divided into two groups after culture with rhGM-CSF, rhIL-4, and rhTNF-α for 9 d. Cells in one group were washed free of cytokines and further cultured for 24 h in the presence of calcium ionophore agent (CI, A23187), while cells in the other group remained in the former culture medium for 24 h. Cells in both of the two groups were analyzed for determining its morphological features, surface antigen expressions, and the ability to stimulate allogeneic mixed lymphocyte reaction (MLR). Results Typical morphological features of DC were found in the cells of both the two groups by inverse microscopy and electron microscopy. High expression levels of CD1α, HLA-DR, CD80, and CD40 were also found in the cells of the both groups. The A23187-treated cells could induce a higher expression of CD86 and CD83, and more proliferating response in allo-MLR compared with the cells which did not treated with A23187. Conclusion The phenotypes and functions of DC generated from PBSC by the treatment of cytokine combination can be further improved by calcium ionophore agent.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2005年第1期44-46,共3页
Immunological Journal
基金
广东省科学技术厅资助项目 (2 0 0 3A30 2 0 30 1 )
