论文部分内容阅读
目的探讨脂质体介导的基因转染对人未成熟树突状细胞(imDC)表型特征及免疫学功能的影响。方法贴壁法分离人脐血来源的单核细胞,用重组人粒细胞巨噬细胞集落刺激因子和重组人白细胞介素4将其诱导分化为imDC后,进行形态学观察和免疫表型鉴定,并将其分为转染组和对照组。转染组将pEGFP-N1质粒通过脂质体转染imDC；对照组不作特殊处理。流式细胞仪检测此法的转染率和两组细胞表面分子[CD86、CD83、人类自细胞DR抗原(HLA-DR)等]的表达率；混合淋巴细胞反应(MLR)检测imDC在转染前后刺激未致敏T淋巴细胞增殖的能力。结果人脐皿来源的imDC形态结构和表面标志符合文献报道的典型特征。脂质体介导的基因转染法转染率约在10%左右。转染组CD86、CD83和HLA-DR的表达率分别为(12±6)%、(8．6±2．3)%和(71±7)%；对照组分别为(13±6)%、(9．1±3．8)%和(72±8)%,两组比较,差异无统计学意义(P＞0．05)。MLR提示imDC转染后其免疫刺激功能较转染前无明显变化(P＞0．05),刺激指数＜2。结论脂质体介导转染imDC后不影响其成熟特性,但转染率不高。
Objective To investigate the effect of liposome-mediated gene transfection on phenotypic characteristics and immunological function of human immature dendritic cells (imDC). Methods Human umbilical cord blood mononuclear cells were isolated by adherence method and then induced to differentiate into imDC by recombinant human granulocyte macrophage colony stimulating factor and recombinant human interleukin-4. Morphological observation and immunophenotyping were performed. And divided into transfection group and control group. Transfection group transfected pEGFP-N1 plasmid by liposome imDC; control group without special treatment. Flow cytometry was used to detect the transfection efficiency of this method and the expression rates of two cell surface molecules [CD86, CD83, human self-cell DR antigen, etc.]; mixed lymphocyte reaction (MLR) Before and after stimulation of non-sensitized T lymphocyte proliferation ability. Results The morphological structure and surface marker of imDC derived from human umbilical dish accorded with the typical features reported in the literature. Liposome-mediated transfection of gene transfection rate of about 10%. The expression rates of CD86, CD83 and HLA-DR in the transfected group were (12 ± 6)%, (8.6 ± 2.3)% and (71 ± 7)%, respectively. The control group were 13 ± 6% , (9.1 ± 3.8)% and (72 ± 8)%, respectively. There was no significant difference between the two groups (P> 0.05). The results of MLR showed that the immunostimulatory function of imDC did not change significantly before transfection (P> 0.05), and the stimulation index was less than 2. Conclusion Liposome-mediated transfection of imDC did not affect its maturation characteristics, but the transfection rate was not high.