TGF-β1受体阻滞剂对树突状细胞融合疫苗制备影响研究.docVIP

TGF-β1受体阻滞剂对树突状细胞融合疫苗制备影响研究 　　[摘要] 目的 探讨TGF-β1受体阻滞剂SB-431542对树突状细胞(DC)融合疫苗制备的影响。方法 ①SB-431542(TGF-β1受体阻滞剂)与粒细胞集落刺激因子(GM-CSF)、白细胞介素-4(IL-4)、脂多糖(LPS)联合应用,刺激C57BL/6小鼠骨髓来源DC成熟,应用聚乙二醇(PEG)融合技术融合DC和小鼠胰腺癌细胞(Panc02),制备融合疫苗。②观察细胞形态、细胞表面表达、MTT法检测肿瘤细胞体外杀伤细胞毒活性。结果 ①在SB-431542参与下,成熟DC的细胞数目及形态无显著改变;②DC表面CD80、CD86、CD40的表达较未加SB-431542显著增高;MTT比色法,添加SB-431542的DC疫苗组对肿瘤细胞的杀伤抑制率显著提高。结论 SB-431542组融合细胞的分子表达较非SB-431542组显著提高;体外MTT显示SB-431542组有更好的抗肿瘤效应,说明SB-431542对增强融合疫苗抗肿瘤效应有明显的增强作用。 　　[关键词] 树突状细胞;细胞融合;SB-431542;肿瘤疫苗 　　[中图分类号] R392.33 [文献标识码] A[文章编号] 1673-9701(2010)14-17-03 　　 　　Effect of TGF-β1Receptor Blocker on Fusion Vaccine of Dendritic Cells 　　REN Jianlin1XU Jun2 　　1. Shanxi Medical University,Taiyuan 030001,China;2. Department of General Surgery,Shanxi Provincial Peoples Hospital,Taiyuan 030012,China 　　 　　[Abstract] Objective To investigate the effect of SB-431542 on the preparation of the dendritic cell fusion vaccine. Methods ①In the presence of SB-431542(an inhibitor TGF-β1 receptor),GM-CSF,IL-4 and LPS,the maturation of dendritic cells from C57BL/6 mouse bone marrow was stimulated,and the preparation of the fusion vaccine was made by use of polyethylene glycol(PEG) fusion technique for fusion of DC and mouse pancreatic cells(Panc02). ②The observation of cell morphology and expression of cell surface phenotypes and the MTT assay detection of specific cytotoxic Tlymphocyte(CTL) were carried out. Results ①SB-431542 could stimulate the full maturation of dendritic cells. Dendritic cells and panc02 cells could be fused by PEG. ②The fusion cells exhibited typical biological characteristics. The expression of cell surface phenotypes was enhanced efficiently. Conclusion SB-431542 can enhance the maturation of dendritic cell. The fusion vaccine could induce a specific and effective immune response against tumor cell. 　　[Key words] Dendritic cells;Cell fusion;SB-431542;Cancer vaccine 　　绝大多数肿瘤缺乏特异性抗原,而且肿瘤组织中抗原提呈细胞数量减少、功能低下或缺如。因此应用细胞融合技术将肿瘤细胞和DC融合,融合细胞即包含肿瘤细胞表面的所有抗原,又具有DC的强大的抗原提呈