痘苗病毒载体介导爱泼斯坦-巴尔病毒潜伏期膜蛋白2A基因转染树突状细胞对其功能的影响

目的：探讨痘苗病毒载体介导爱泼斯坦－巴尔病毒（EBV）潜伏期膜蛋白2A（LMP2A）基因转染树突状细胞（DC）对DC的表型和功能的影响，以及EBV相关肿瘤免疫治疗性疫苗的可行性，方法：用EBV－LMP2A基因重组痘苗病毒（Vac-LMP2A)转染成熟的DC，流式细胞术（FACS）检测转染前后DC表面分子CD1a,CD83,CD40,CD80,HLA-DR变化，3H－TdR掺入法检测转染前后DC刺激同种异体T淋巴细胞增殖等功能的改变。结果：转染后LMP2A蛋白在DC细胞内高表达，Vac-LMP2A转染成熟DC前后对其表面共刺激分子及特征性表面标志无影响，转染后的DC仍具有较强的刺激同种异体TI林巴细胞增殖的能力。结论：痘苗病毒载体是介导外源基因LMP2A转染DC的有效载体，Vac-LMP2A转染成熟DC对DC表型和功能无明显影响，是EBV相关肿瘤如鼻咽癌免疫治疗的理想载体。

Study on Effects on Function of Dendritic Cells Transfected with the Gene of Epstein- Barr Virus Latent Membrane Protein 2A Mediated by Recombinant Vaccinia Virus

gene,and provide further evidence for investigation on the therapeutic vaccine against EBV-associated malignancies. Methods:DC were transfected with EBV-LMP2A recombinant vaccinia virus (Vac-LMP2A). Before and after the transfection,the expression of surface markers on mature DC including CD1a,CD83,CD40,CD80,HLA DR were compared by means of FACS and the function of mature DC to stimulate allogenetic T cells reaction was measured by methods of mixed leukocyte reaction (MLR) and 3H thymidine (3H TdR) uptake test. Results:There were on significant changes in the surface markers expression and in the 3H TdR uptake test before and after the transfection. Transfected DC still had strong potential to stimulate the proliferation of allogenetic T cells. Conclusion:Vaccinia virus vector is safe to mediate the transfection of LMP2A into DC. The function of mature DC was not significantly affected by Vac LMP2A transfecting. This study could pave the way for the further treatment of EBV associated malignancies.