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目的观察甲状腺相关眼病(thyroid-associated ophthal mopathy,TAO)患者眼眶结缔组织中共刺激分子CD40/CD40L的表达,并探讨其可能的作用机制。方法采用免疫组化检测CD40/CD40L、细胞间黏附分子1(intercellular adhesionmolecule1,ICAM-1)/白细胞功能抗原(1eukocyte function antigen1,LFA-1)在甲状腺相关眼病患者(n=12)及正常对照(n=8)眼眶组织中的表达情况;原代培养TAO患者及正常对照眼眶成纤维细胞,应用RT-PCR方法检测CD40L刺激后眼眶成纤维细胞合成ICAM-1mRNA水平的变化。结果 TAO患者眼眶结缔组织中可见高表达CD40、ICAM-1的成纤维细胞,大量淋巴细胞浸润并高表达LFA-1;双酶标组化显示CD40阳性表达的成纤维周围浸润CD40L阳性表达的淋巴细胞;ICAM-1阳性眼眶成纤维细胞与LFA-1阳性淋巴细胞以直接接触的形式存在。RT-PCR结果显示,未激活状态TAO组眼眶成纤维细胞ICAM-1mRNA水平高于正常组(P<0.05);两组眼眶成纤维细胞经CD40L诱导后ICAM-1mRNA水平高于未激活状态(P<0.05);ICAM-1mRNA表达量与CD40L刺激呈剂量依赖关系。结论 CD40/CD40L共刺激分子在TAO患者眼眶结缔组织中高表达,可能在TAO眼眶淋巴细胞浸润及炎症发生、发展中起重要作用。
Objective To investigate the expression of costimulatory molecule CD40 / CD40L in orbital connective tissue in patients with thyroid-associated ophthalmopathy (TAO) and to explore its possible mechanism. Methods The expressions of CD40 / CD40L, ICAM-1 / LFA-1 were detected by immunohistochemistry in patients with thyroid associated ophthalmopathy (n = 12) and normal controls n = 8) orbital tissue; primary cultured TAO patients and normal control orbital fibroblasts, using RT-PCR method to detect CD40L stimulated orbital fibroblasts synthesis of ICAM-1 mRNA levels. Results In TAO patients, high expression of CD40 and ICAM-1 fibroblasts were found in the orbital connective tissues, with a large number of lymphocytes infiltrating and high expression of LFA-1. The double-labeled biopsy showed CD40-positive fibroblast infiltrating CD40L positive lymphatic Cells; ICAM-1-positive orbital fibroblasts exist in direct contact with LFA-1-positive lymphocytes. The results of RT-PCR showed that the level of ICAM-1 mRNA in orbital fibroblasts in inactive TAO group was higher than that in normal group (P <0.05). The ICAM-1 mRNA level in orbital fibroblasts induced by CD40L was higher than that in inactive group <0.05). The expression of ICAM-1mRNA and CD40L stimulated in a dose-dependent manner. Conclusions CD40 / CD40L costimulatory molecules are highly expressed in orbital connective tissue of patients with TAO, which may play an important role in the infiltration of TAO and the occurrence and development of inflammation in TAO patients.