Interleukin-6 Production in CD40-Engaged Fibrocytes in Thyroid-Associated Ophthalmopathy: Involvement of Akt and NF-κB
Interleukin-6 Production in CD40-Engaged Fibrocytes in Thyroid-Associated Ophthalmopathy: Involvement of Akt and NF-κB
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DOI：
10.1167/iovs.12-9861
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发表时间：
2012-11-01
影响因子：
4.4
通讯作者：
Douglas, Raymond S.
中科院分区：
文献类型：
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作者：
Gillespie, Erin F.;Raychaudhuri, Nupur;Douglas, Raymond S.
目的。 CD40-CD40 配体 (CD40L) 相互作用似乎在自身免疫性疾病中发挥致病作用。在这里，我们量化了纤维细胞、循环细胞和骨髓来源的祖细胞上的 CD40 表达。 CD40结扎的功能性后果是确定的，因为它们可能促进与甲状腺相关性眼病（TAO）相关的组织重塑。方法。通过流式细胞术测定格雷夫斯病 (GD) 患者培养的纤维细胞和眼眶成纤维细胞 (GOFB) 上的 CD40 水平以及纤维细胞丰度。通过实时 PCR 评估 CD40 mRNA 表达，而通过 Luminex 和 RT-PCR 测量对 CD40 连接的反应。通过蛋白质印迹和免疫荧光测定蛋白激酶 B (Akt) 和核因子 (NF)-kappa B (NF-kappa B) 信号传导。 结果。纤维细胞上的基础 CD40 表达高于 GOFB 上的表达。 IFN-γ 上调两种细胞类型中的 CD40，其作用是在翻译前水平介导的。纤维细胞响应 CD40L 产生高水平的细胞因子，包括白细胞介素 6 (IL-6)、TNF-α、IL-8、MCP-1 和 RANTES（调节正常 T 细胞表达和分泌的激活）。 IL-6 诱导是稳态 IL-6 mRNA 增加的结果，并通过 Akt 和 NF-κ B 激活介导。与健康受试者相比，TAO 供体体内循环 CD40(+)CD45(+)Col1(+) 纤维细胞的出现频率要高得多。结论。纤维细胞显示出特别高水平的功能性 CD40。 CD40L 引发的信号传导导致多种细胞因子的产生。其中，IL-6 表达通过 Akt 和 NF-kappa B 途径介导。 TAO患者中循环CD40(+)纤维细胞的频率显着增加，表明该受体可能代表TAO的治疗靶点。 （投资眼科可见科学。2012 年；53：7746-7753）DOI：10.1167/iovs.12-9861
PURPOSE. CD40-CD40 ligand (CD40L) interactions appear to play pathogenic roles in autoimmune disease. Here we quantify CD40 expression on fibrocytes, circulating, and bone marrow-derived progenitor cells. The functional consequences of CD40 ligation are determined since these may promote tissue remodeling linked with thyroid-associated ophthalmopathy (TAO).METHODS. CD40 levels on cultivated fibrocytes and orbital fibroblasts (GOFB) from patients with Graves' disease (GD), as well as fibrocyte abundance, were determined by flow cytometry. CD40 mRNA expression was evaluated by real-time PCR, whereas response to CD40 ligation was measured by Luminex and RT-PCR. Protein kinase B (Akt) and nuclear factor (NF)-kappa B (NF-kappa B) signaling were determined by Western blot and immunofluorescence.RESULTS. Basal CD40 expression on fibrocytes is greater than that on GOFB. IFN-gamma upregulates CD40 in both cell types and its actions are mediated at the pretranslational level. Fibrocytes produce high levels of cytokines, including interleukin-6 (IL-6), TNF-alpha, IL-8, MCP-1, and RANTES (Regulated on Activation, Normal T Cell Expressed and Secreted) in response to CD40L. IL-6 induction results from an increase in steady state IL-6 mRNA, and is mediated through Akt and NF-kappa B activation. Circulating CD40(+)CD45(+)Col1(+) fibrocytes are far more frequent in vivo in donors with TAO compared with healthy subjects.CONCLUSIONS. Particularly high levels of functional CD40 are displayed by fibrocytes. CD40L-provoked signaling results in the production of several cytokines. Among these, IL-6 expression is mediated through Akt and NF-kappa B pathways. The frequency of circulating CD40(+) fibrocytes is markedly increased in patients with TAO, suggesting that this receptor might represent a therapeutic target for TAO. (Invest Ophthalmol Vis Sci. 2012;53:7746-7753) DOI:10.1167/iovs.12-9861