CMVpp65基因修饰的树突状细胞刺激自身T细胞活化  

CMV pp65 Gene Modified Dendritic Cells Activate Autologous T Cells

作　　者：高广勋[1] 陈协群[1] 张劲翼[2] 朱华锋[1] 董宝侠[1] 顾宏涛[1] 高瑛[1] 潘耀柱[1] 

机构地区：[1]第四军医大学西京医院血液科,陕西西安710032 [2]多伦多大学西奈山医院Samuel Leunenfeld研究所,加拿大

出　　处：《中国实验血液学杂志》2008年第2期397-400,共4页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目,编号30370599

摘　　要：巨细胞病毒(CMV)感染易发生于慢性移植物抗宿主病(cGVHD)患者,其特异性免疫依赖于T细胞活性。本研究探讨CMV pp65基因修饰的树突状细胞(DC)对自身T细胞的活化作用。采用具有高效转染非分裂细胞的慢病毒系统将CMV全长pp65基因导入鼠源性DC;采用CpG-DNA诱导DC细胞成熟;采用流式细胞术及免疫荧光方法测定T淋巴细胞抗原及IFNγ表达。结果表明:慢病毒感染DC的感染复数(multiplicity of infection,MOI)为50,最佳感染效率可达30%-40%;表达pp65基因的成熟DC能刺激自身naiveT细胞表达CD69;表达PP65蛋白的成熟DC能够刺激自身CD4+或CD8+T细胞分泌IFNγ。结论:pp65基因修饰、CpG-DNA诱导成熟的DC能体外激活CMV特异性T淋巴细胞。Cytomegalovirus （CMV） infection is a dangerous complication in patients with chronic graft versus host disease （cGVHD）. CMV-specific immunity depends on the activity of T cells. This study was aimed to investigate the effect of CMV pp65 gene modified dendritic cells （DCs） on activation of autologous T cells. Lentivirus system was utilized to introduce the CMV full-length pp65 gene into mouse DCs; CpG-DNA was used to induce mature DCs; flow cytometry and immunofluorescence were used to determine the expression of antigen and IFNγ in T lymphocytes. The results showed that the DCs were infected with lentivirus at a multiplicity of infection （ MOI ） of 50 with optimal infectious efficiency of 30% - 40% ; mature DCs expressing pp65 gene could stimulate autologous naive T cells to express CD69 specifically ; mature DCs expressing PP65 could stimulate autologous CD4^＋ or CD8^＋ T cells to produce IFNγ. It is concluded that CMV pp65-modified and CpG-DNA-induced mature DCs can activate CMV- specific T lymphocytes in vitro.

关 键 词：巨细胞病毒 PP65蛋白 T淋巴细胞 树突状细胞 CpG—DNA 

分 类 号：Q813[生物学—生物工程] R392[医药卫生—免疫学]