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

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.