人DeltaNp73α基因修饰的树突状细胞抗瘤免疫学效应研究  

In vitro anti-tumor immune response induced by dendritic cells transfected with DeltaNp73α recombinant adenovirus

作　　者：胡义杰[1] 范士志[1] 蒋耀光[1] 李志平[1] 陈建明[1] 何勇[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所全军胸外科中心,重庆400042

出　　处：《第三军医大学学报》2007年第9期763-766,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30600611);第三军医大学中青年基金(XG200540)~~

摘　　要：目的研究DeltaNp73α重组腺病毒转染树突状细胞诱导的特异性免疫抗瘤作用。方法人脐静脉血经梯度离心及IL-4、GM-CSF诱导培养树突状细胞;DeltaNp73α重组腺病毒增强离心法转染树突状细胞,观察DeltaNp73α对树突状细胞活化、凋亡的影响;同时检测其所诱导的DeltaNp73α特异性细胞毒性T淋巴细胞对不同DeltaNp73α表达水平肿瘤细胞的细胞毒性作用。结果Western blot证实重组腺病毒转染树突状细胞后DeltaNp73α的有效表达;DeltaNp73α修饰的树突状细胞凋亡率显著低于对照组;DeltaNp73α修饰树突状细胞诱导的细胞毒性T细胞可以有效的杀伤DeltaNp73α高表达的A549/DeltaNp73α细胞、K-562细胞;且这种免疫抗瘤作用可能与靶肿瘤细胞DeltaNp73α水平相关。结论DeltaNp73α修饰的树突状细胞,其凋亡率降低;并可以有效诱导针对DeltaNp73α高表达肿瘤细胞的特异免疫抗瘤作用。Objective To investigate the effects of dendritic cells infected with adenovirus vector encoding DeltaNp73α on breaking the immune tolerance and inducing immunity against DeltaNp73α-overexpressing tumor. Methods Immature dendritic cells generated in the presence of interleukin-4 and granulocyte/macrophage colony-stimulating factor from human umbilical cord blood were transfected with recombinant adenovirus vector encoding full-length human DeltaNp73α cDNA by centrifugal force method. Viability and activation status of DCs after transfection were assessed by flow cytometric analysis, and induction of DeltaNp73α-specific CTL response was also evaluated by the cytotoxic assay against DeltaNp73α-overexpressing human tumor cells. Results DeltaNp73α expression in the transfected DCs was confirmed by Western blotting. DeltaNp73α enhanced DCs survival. A significant DeltaNp73α-specific CTL-stimulatory ability were observed in the DehaNp73α-overexpressing cell lines, A549/DeltaNp73α and K-562. Conclusion DeltaNp73α enhances the survival of dendritic cells. Adenovirus vector-mediated DeltaNp73α-transfected DCs can effectively induce strong antigen-specific T cell response.

关 键 词：DehaNp73α 树突状细胞 重组腺病毒 免疫抗瘤 

分 类 号：R392.5[医药卫生—免疫学] R394[医药卫生—基础医学]