Goldberg Michael, Nadiv Orna, Luknar-Gabor Noemi, Agar Gabriel, Beer Yiftah, Katz Yitzhak
Allergy and Immunology Institute,Assaf-Harofeh Medical Center, Zerifin 70300, Israel.
Mol Immunol. 2009 May;46(8-9):1854-9. doi: 10.1016/j.molimm.2009.01.004. Epub 2009 Feb 6.
In order to determine the mechanisms by which a chronic inflammatory network can be maintained in the arthritic joint, we examined whether fibroblast-like synoviocytes (FLS) could provide feedback signals after their stimulation by inflammatory cytokines. FLS and dermal fibroblasts (DF) were derived from rheumatoid arthritis (RA), osteoarthritis (OA) and post-trauma patients. These two cell types were then stimulated with 10 nanogram/ml of TNFalpha, IL-1beta and IL-17 alone or in combination treatments. Specific mRNA expression of IL-23 p19 was quantitated by real-time PCR and its protein by immunoprecipitation. A striking specific synergistic induction of IL-23 p19 versus IL-12 p35 mRNA expression was noted after stimulation with IL-17 and TNFalpha in FLS, and to a lesser degree in DF (p<0.043). This synergistic response was composed of an initial priming step by IL-17, thus making FLS hyperresponsive to TNFalpha-mediated stimulation. In contrast, IL-1beta mediated induction of IL-23 p19 expression was cell-specific. Induction of IL-23 p19 expression by IL-1beta was present in FLS but almost absent in the DF derived from the same patients. Furthermore, IL-1beta did not synergize with IL-17 to induce IL-23 p19 expression. Immunoprecipitation of FLS cellular lysates after stimulation with IL-17 and TNFalpha detected p19 protein and this was enhanced by the addition of IL-1beta. However, no co-immunoprecipitation of the p40 subunit of IL-23 was noted from the same cells. Thus, FLS are potently regulated by inflammatory cytokines to specifically express IL-23 p19. Additional byproducts of the inflammatory milieu may be required for the generation and secretion of bioactive IL-23.
为了确定慢性炎症网络在关节炎关节中得以维持的机制，我们研究了成纤维样滑膜细胞（FLS）在受到炎性细胞因子刺激后是否能够提供反馈信号。FLS和真皮成纤维细胞（DF）分别取自类风湿性关节炎（RA）、骨关节炎（OA）和创伤后患者。然后，用10纳克/毫升的肿瘤坏死因子α（TNFα）、白细胞介素-1β（IL-1β）和白细胞介素-17（IL-17）单独或联合处理这两种细胞类型。通过实时聚合酶链反应（PCR）对IL-23 p19的特异性信使核糖核酸（mRNA）表达进行定量，并通过免疫沉淀法对其蛋白质进行定量。在用IL-17和TNFα刺激后，在FLS中观察到IL-23 p19与IL-12 p35 mRNA表达有显著的特异性协同诱导作用，在DF中这种作用程度较小（p<0.043）。这种协同反应由IL-17引发的初始致敏步骤组成，从而使FLS对TNFα介导的刺激反应增强。相比之下，IL-1β介导的IL-23 p19表达诱导具有细胞特异性。IL-1β诱导IL-23 p19表达在FLS中存在，但在来自同一患者的DF中几乎不存在。此外，IL-1β与IL-17不协同诱导IL-23 p19表达。用IL-17和TNFα刺激后，对FLS细胞裂解物进行免疫沉淀检测到p19蛋白，添加IL-1β可增强这种蛋白的表达。然而，在相同细胞中未观察到IL-23的p亚基40的共免疫沉淀。因此，FLS受到炎性细胞因子的有效调节以特异性表达IL-23 p19。炎性环境的其他副产物可能是生物活性IL-23产生和分泌所必需的。
