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目的研究HBV无症状携带者树突状细胞的分化及功能。方法抽取健康对照者(A组)、HBV无症状携带者(B组)外周静脉血,经密度梯度离心法分离获得PBMC,用TNF-αI、L-10分别和rhIL-4r、hGM-CSF刺激培养,获得:成熟树突状细胞(mDC)、不成熟树突状细胞(imDC)。rhIL-4r、hGM-CSF刺激培养获得树突状细胞(DC)作组内对照。imDC组加入TNF-α刺激,检验imDC的稳定性。流式细胞仪检测各组反映树突状细胞成熟度的各种分子。结果A、B组内mDC细胞膜表达HLA-DR,HLA-ABC分子,分泌的IL-12p70与imDC、DC、imDC+TNF-α组相比明显增加;A、B组间mDC的HLA-DR,HLA-ABC表达无显著性差异,IL-12p70分泌差异有统计学意义(P<0.01)。imDC的HLA-ABC、HLA-DR低表达,IL-12p70分泌也较低,与本组内DC比较结果差异有统计学意义(P<0.01);A、B组间imDC的HLA-DR,HLA-ABC低表达,IL-12p70分泌差异无统计学意义。imDC加入TNF-α后HLA-ABC、HLA-DR仍低表达,IL-12p70分泌低于本组内DC,结果差异有统计学意义(P<0.01);A、B组间结果无显著性差异。结论在A、B组,TNF-α均有效促进DC成熟为mDC,IL-10均抑制DC的成熟,不受A、B组细胞来源影响。mDC分泌大量IL-12p70,并在B组低于A组,说明B组mDC存在一定的功能不足。
Objective To study the differentiation and function of dendritic cells in asymptomatic HBV carriers. Methods Peripheral venous blood was collected from healthy controls (group A) and HBV asymptomatic carriers (group B). PBMC were isolated by density gradient centrifugation and stimulated with TNF-αI, L-10 and rhIL-4r, hGM-CSF Cultivation, access: mature dendritic cells (mDC), immature dendritic cells (imDC). rhIL-4r, hGM-CSF stimulated to obtain dendritic cells (DC) as an in-group control. imDC group added TNF-α stimulation, test imDC stability. Flow cytometry was used to detect various molecules that reflected the maturation of dendritic cells. Results The expression of HLA-DR and HLA-ABC molecules in the membrane of mDC in group A and group B was significantly increased compared with those in imDC, DC and imDC + TNF-α group. The levels of HLA-DR, HLA-ABC expression was no significant difference, IL-12p70 secretion difference was statistically significant (P <0.01). The expression of HLA-ABC, HLA-DR and IL-12p70 in imDC were also lower than that in imDC (P <0.01) -ABC low expression, IL-12p70 secretion difference was not statistically significant. The expression of HLA-ABC and HLA-DR in imDC was still low, while the secretion of IL-12p70 was lower than that in this group (P <0.01). There was no significant difference between groups A and B . Conclusion Both A and B groups and TNF-α can effectively promote the maturation of DCs to mDC. Both IL-10 and IL-10 inhibited the maturation of DCs, and were not influenced by the cell sources of A and B groups. mDC secreted a large amount of IL-12p70 and was lower in group B than in group A, indicating that mDC in group B had some insufficiency.