反义bcl-2的表达诱导癌细胞凋亡致敏树突状细胞的免疫应答

目的 观察树突状细胞（DCs)对反义bcl-2的逆转录病毒表达载体诱导的凋亡胆管癌细胞获取抗原后，抗肿瘤免疫应答及对胆管癌细胞的特异性免疫杀伤效果。方法 用粒－巨噬细胞集落刺激因子（GM－CSF）加白介素－4（IL－4）从人外周血分化，诱导DCs，反义bcl-2的逆转录病毒表达载体在体外诱导培养的人胆管癌细胞凋亡，将DCs，T淋巴细胞和凋亡胆管癌细胞区培养，同时设计不同类型肿瘤细胞（坏死胆管癌细胞及培养胆管癌细胞）作对照，7d后，分离，富集DCs，T淋巴细胞进行免疫应答及肿瘤细胞杀伤试验。结果 DCs高表达共刺激分子B7和CD1a，表面具有典型不规则突起，反义bcl-2表达诱导癌细胞凋亡形成的凋亡小体被DCs捕捉，吞噬，负载抗原的DCs可以有效提呈胆管癌细胞抗原，有强烈的免疫应答，刺激的细胞毒T淋巴细胞（CTLs)特异性地杀伤胆管癌细胞。结论 反义bcl-2的逆转录病毒表达载体诱导癌细胞凋亡可以致敏rhGM-CSF加rhIL-4从人外周血单核细胞诱导，扩增出的DCs并产生显著的杀伤胆管癌细胞的免疫反应，可望成为特异性免疫治疗肿瘤的一条新途径。

Immune responses of dendritic cells acquiring antigen from apoptot ic hepatochlangioma cells caused by antisense bcl-2 expression

Objective To investigate the induction CTLs of antitumor response and therapeutic effects after DCs acquire antigen from apoptotic hepatochlangioma cells caused by antisense bcl 2 expression sector.Methods DCs from blood mononuclear that maintain the antigen capturing and processing capacity charateristic of immature have been established in vitro by using granulocyte/macrophage colony stimulating factor(GM CSF) and interleukin 4.Then, hepatochlangioma cells were induced apoptosis with antisense bcl 2 expression sector.We design these experiment groups including (1)coculture of DCs and apoptotic cancer cells and T cells,(2)coculture of DCs and necrosis cancer cells and T cells,(3)coculture of DCs and cultured cancer cells and T cells.They are cocultured for 7 days.DCs and T cells are riched separately.Finally, antitumor response test have been done.Results These cells have typical dendritic morphology, express high levels of CD1a, BT and acquire antigen from apoptotic cells caused by antisense bcl 2 expression and obviously induce on T cell stimulatory capacity in MLR.Conclusion We have established DCs from blood mononuclear using GM CSF and IL 4 and DCs can efficiently prensent antigen drived from apoptotic cells caused by antisense bcl 2 expression sector and induce T cells increasing obviously.It probably become effective approach of DCs transduced with antigen.