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为研究瓜氨酸化波形蛋白(citrullinated vimentin,cVim)对外周血单个核细胞来源树突状细胞(dendritic cells,DC)的成熟和功能的影响,本文通过分离外周血单个核细胞,GM-CSF和IL-4体外培养制备未成熟DC(immature DC,imDC),分别用脂多糖(LPS)、波形蛋白(vimentin,Vim)和cVim刺激培养的imDC,流式细胞仪检测DC表面标志CD14、CD80、CD83、CD86、MHCⅠ和MHCⅡ表达变化,ELISA方法检测培养上清液IL-6、IL-10、IL-12、IL-23、和IFN-γ含量。LPS显著诱导DC表面MHCⅡ、CD83、CD80和CD86表达上调(P值均<0.05);cVim(10μg/ml)诱导imDC表达CD80和MHCⅡ增加(P<0.05),并促进IL-10、IL-12和IFN-γ分泌(P<0.01);而Vim(10μg/ml)仅促进imDC表达MHCⅡ(P<0.01)和分泌IL-10(P<0.01),并抑制IL-23表达(P<0.01)。本研究表明Vim经瓜氨酸化修饰后有促进外周血单个核细胞来源imDC成熟、分泌炎症性细胞因子的作用。
In order to investigate the effect of citrullinated vimentin (cVim) on the maturation and function of dendritic cells (DCs) derived from peripheral blood mononuclear cells, IL-4 were cultured in immature DC (imDC) in vitro. The cultured imDC were stimulated with lipopolysaccharide (LPS), vimentin (Vim) and cVim respectively. The surface markers CD14, CD80, CD83, CD86, MHC Ⅰ and MHC Ⅱ were detected by ELISA. The contents of IL-6, IL-10, IL-12, IL-23 and IFN- LPS significantly upregulated the expression of MHCⅡ, CD83, CD80 and CD86 on DCs (all P <0.05); cDim (10μg / ml) induced imDC expression of CD80 and increased MHCⅡ (P <0.05) (P <0.01), and inhibited the expression of IL-23 (P <0.01). However, Vim (10μg / ml) promoted the expression of MHCⅡ (P <0.01) and IL- . This study shows that Vim citrullination can promote the maturation of peripheral blood mononuclear cells derived imDC, the secretion of inflammatory cytokines.