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目的观察重组腺病毒载体AdEASY-增强型绿色荧光蛋白(EGFP)转染人未成熟树突状细胞(imDC)后,其表型特征及免疫学功能的变化,并探讨白细胞介素(IL)10对腺病毒转染诱导imDC成熟的抑制作用。方法贴壁法分离人脐带血来源的单核细胞,利用重组粒细胞巨噬细胞集落刺激因子(GM-CSF)和IL-4诱导分化imDC。对照组为常规培养的imDC,转染组用AdEASY-EGFP转染imDC,IL-10组用IL-10处理转染后细胞。流式细胞仪检测细胞表面成熟标志[CD86、CD83和人类白细胞DR抗原(HLA-DR)],混合淋巴细胞反应(MLR)检测其刺激同种异体未致敏T淋巴细胞的增殖能力。结果腺病毒转染imDC后,转染组细胞成熟表型表达率分别为CD86:46±10、CD83: 38±7、HLA-DR:82±10,均较对照组(10±7、8±3、68±8)显著上调,且刺激T淋巴细胞增殖的能力显著加强(SI>2．0)。IL-10组处理的imDC上述表型表达率分别为CD86:8±5、CD83:9±3、HLA-DR: 63±12,与对照组比较差异无统计学意义(P>0．05),其刺激T淋巴细胞增殖的能力也明显下降。结论腺病毒能有效转染imDC,但在转染后有促进其成熟的趋势;用IL-10能有效抑制该成熟状态。
Objective To observe the phenotypic characteristics and immunological function of recombinant adenovirus vector AdEASY-EGFP transfected with human immature dendritic cells (imDC) and to investigate the effects of interleukin (IL) 10 Adenovirus transfection induced imDC maturation inhibition. Methods Human umbilical cord blood mononuclear cells were isolated by adherence method, and then imDC was induced by GM-CSF and IL-4. The control group was routinely cultured imDC, the transfected group transfected with AdEASY-EGFP imDC, IL-10 group treated with IL-10 transfected cells. Flow cytometry was used to detect the maturation markers of cell surface [CD86, CD83 and human leukocyte antigen (HLA-DR)], and mixed lymphocyte reaction (MLR) to detect the proliferation of allogeneic non-sensitized T lymphocytes. Results After imDC transfection, the mature phenotypes of the transfected cells were CD86: 46 ± 10, CD83: 38 ± 7 and HLA-DR: 82 ± 10, respectively, which were significantly higher than that of the control group (10 ± 7, 8 ± 3,68 ± 8), and significantly enhanced the ability of stimulating T lymphocyte proliferation (SI> 2.0). The expression rates of imDC in the group treated with IL-10 were respectively CD86: 8 ± 5, CD83: 9 ± 3 and HLA-DR: 63 ± 12, showing no significant difference from the control group (P> 0.05) , Its ability to stimulate T lymphocyte proliferation also decreased significantly. Conclusions Adenovirus can effectively transfect imDC, but it has the tendency to promote its maturation after transfection; IL-10 can effectively inhibit the mature state.