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目的 体外培养慢性乙型肝炎病人的树突状细胞 ,并从细胞表型和功能方面与正常人树突状细胞进行比较。方法 从慢性乙型肝炎病人及正常人外周血分离单个核细胞 ,加入含 10 0 0U/mlGM CSF和 5 0 0U/mlIL 4的无血清培养基AIM V体外培养 ,获得树突状细胞。利用流式细胞仪检测细胞因子TNF α对树突状细胞培养的影响 ,IL 12ELISA试剂盒检测树突状细胞分泌IL 12的水平 ,并采用MLR方法检测树突状细胞刺激同种异体T淋巴细胞增殖的能力。结果 1.慢性乙型肝炎病人的外周血单个核细胞用AIM V培养及细胞因子诱导后 ,可获得成熟的具有典型形态的树突状细胞 ,加入TNF α后 ,IL 12分泌增高 ,CD83表达增加 ;2 .病人树突状细胞与正常人比较 ,CD80表达较正常人的低 (P <0 .0 5 ) ,刺激同种异体T细胞增殖能力低于正常人。结论 1.慢性乙型肝炎病人的树突状细胞与正常人一样可用AIM V无血清培养基及特定的细胞因子诱导在体外大量获得 ;2 .慢性乙型肝炎病人的树突状细胞与正常人相比较在功能上降低 ,在形态数量上差异无显著意义。
Objective To culture dendritic cells from patients with chronic hepatitis B in vitro and to compare with normal human dendritic cells in terms of cell phenotype and function. Methods Mononuclear cells were isolated from peripheral blood of patients with chronic hepatitis B and normal controls and cultured in serum-free medium AIM V containing 100 μM / ml GM-CSF and 500 μL / ml IL-4 to obtain dendritic cells. The effect of TNFα on the dendritic cell culture was detected by flow cytometry. IL 12 ELISA kit was used to detect the level of IL 12 secreted by dendritic cells. The dendritic cells were stimulated by dendritic cells to stimulate allogeneic T lymphocytes Ability to proliferate. Peripheral blood mononuclear cells from chronic hepatitis B patients induced by AIM V and induced by cytokines could produce mature dendritic cells with typical morphology. After addition of TNFα, IL 12 secretion increased and CD83 expression increased ; 2. Dendritic cells of patients compared with normal people, CD80 expression was lower than normal (P <0. 05), stimulate the proliferation of allogeneic T cells lower than normal. Dendritic cells from patients with chronic hepatitis B can be obtained in large quantities in vitro with AIM V-free serum-free medium and specific cytokines as well as normal human; 2. Dendritic cells from patients with chronic hepatitis B are significantly different from normal people Compared with the function of lower, the number of morphological differences was not significant.