HCV C-Fc基因转染的小鼠树突状细胞促进混合淋巴细胞反应的作用

Acceleration of mixed lymphocyte reaction by HCV C-Fc gene-transferred murine dendritic cells

目的 :观察HCVC Fc基因电穿孔法转染的小鼠树突状细胞(DC)功能的改变。方法 :分离小鼠骨髓单个核细胞与rmGM CSF和rmIL 4培养 1wk,对培养的细胞进行形态学观察 ,并用FACS检测细胞表面DEC2 0 5的表达。以通过质粒pcD NA3HCVC Fc电穿孔法转染培养的DC ,用间接免疫荧光染色测定转染细胞中HCVC Fc的水平 ;用混合淋巴细胞反应检测电穿孔法转染细胞的功能。结果 :培养 1wk后 ,得到具有典型DC形态的细胞 ,以质粒pcDNA3HCVC Fc为载体电穿孔法转染DC后 ,转染细胞中可检测到较高水平的HCVC Fc。与对照组相比较以电穿孔法转染的细胞能明显刺激混合淋巴细胞反应。结论 :小鼠骨髓单个核细胞加GM CSF和IL 4培养 1wk ,可生成大量的DC。质粒pcDNA3HCVC Fc转染培养的DC对T细胞刺激作用显著增强。

AIM: To observe the metergasis of murine dendritic cells (DCs) transfected with HCV C Fc gene through electroporation. METHODS: Mononucleocytes isolated from murine bone marrow were co cultured with rmGM CSF and rm IL 4 for 7 days. Morphological characteristics of the cultured cells were observed under scan electron microscope (SEM) and the expression of DEC205 on the cells was detected by FACS. DCs derived from the culture were transfected with plasmids containing HCV C Fc gene. HCV C Fc level in the transfected cells was detected by indirect immunofluorescence assay. MLR was studied with DCs and T cells. RESULTS: Following 7 day culture, a large number of cells with typical characteristics of DC were observed. The HCV C Fc level in the transfected DCs was higher. MLR was stimulated markedly by DCs transfected with HCV C Fc gene in comparison with the control group. CONCLUSION: A large number of DCs could be generated from murine bone marrow mononucleocyte cultures supplemented with GM CSF and IL 4 for 1 week. The function of DCs transfected with pcDNA3HCV C Fc was enhanced in MLR.