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目的 研究转化生长因子 β1(TGF β1)对树突细胞 (dendriticcells,DC)功能的影响。方法 在培养体系中应用不同的细胞因子培养未成熟DC (imDC ,GM CSF)和TGF β1处理的DC(TGFβ DC ,GM CSF +TGF β1) ,观察其对脂多糖 (LPS)刺激的反应。透射电镜观察细胞超微结构 ,流式细胞仪检测细胞表型 ,BrdUELISA法检测DC刺激异基因T细胞增殖的能力 ,ELISA法检测DC在LPS刺激后分泌IL 12 p70的水平 ,逆转录 聚合酶链反应 (RT PCR)方法检测Toll like受体 4(TLR4 )表达。结果 与imDC相比 ,TGFβ DC在LPS刺激后仍能保持未成熟的细胞形态。TGFβ DC的CD80 ,CD86表达明显低于imDC[(4 .14± 0 .95 ) %和 (13.90± 7.2 2 ) % ;(8.6 0± 0 .75 ) %和 (2 0 .6 3± 5 .0 3) % ,P值均 <0 .0 5 ]。ImDC对LPS有更强的反应性 ,其中I Ab、CD80升高的幅度明显高于TGFβ DC(P值分别 <0 .0 1及 <0 .0 5 )。TGFβ DC在 96h的混合淋巴细胞反应中 ,DC/T细胞为 1∶4 ,1∶1时 ,TGFβ DC的异基因刺激能力较imDC弱 (P值均 <0 .0 5 )。LPS刺激TGFβ DC 2 4h后分泌IL 12p70的能力显著低于imDC(P <0 .0 1) ,TGFβ DC较imDC弱表达TLR4 (P <0 .0 5 )。 结论 TGFβ能抑制DC共刺激分子的表达 ,且能抵抗LPS的促成熟作用 ,并可能与其TLR4的表达下降?
Objective To investigate the effect of transforming growth factor-β1 (TGF-β1) on the function of dendritic cells (DCs). Methods Different DCs were cultured in immature DCs (imDC, GM CSF) and TGFβ1-treated DCs (TGFβ DC, GM CSF and TGF β1) in culture system to observe their response to lipopolysaccharide (LPS) stimulation. The ultrastructure of the cells was observed by transmission electron microscopy, the cell phenotype was detected by flow cytometry, the ability of DCs to stimulate the proliferation of allogeneic T cells was detected by BrdU ELISA, the level of IL 12 p70 secreted by DCs was measured by ELISA, Toll like receptor 4 (TLR4) expression was detected by RT PCR. Results Compared with imDC, TGFβ DC still retained immature cell morphology after LPS stimulation. The expression of CD80 and CD86 in TGFβ DC was significantly lower than those in imDC [(4.14 ± 0.95)% and (13.90 ± 7.22)%; (8.6 ± 0.75)% and (2.06 ± 3.5)% respectively. 0 3)%, all P <0. 05]. ImDC was more responsive to LPS, with a significant increase in I Ab and CD80 over TGFβ DC (P values <0.01 and <0.05, respectively). In the mixed lymphocyte reaction of TGFβ DC at 96h, the ability of allogeneic stimulation of TGFβ DC was weaker than that of imDC when DC / T cells were 1: 4, 1: 1 (all P <0.05). The ability of LPS to stimulate IL-12p70 secretion by TGFβ DC 2 for 4 h was significantly lower than that of imDC (P <0.01). TGFβ DC expressed TLR4 weakly than imDC (P <0.05). Conclusion TGFβ can inhibit the expression of DC costimulatory molecules and resist the pro-mature effect of LPS, and may be related to the decrease of TLR4 expression.