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通过研究干燥综合征(Sjgren’s syndrome,SS)患者与健康对照组唾液腺上皮细胞CD40、ICAM-1/CD54蛋白及基因表达水平的差异,揭示SS患者唾液腺上皮细胞异常免疫活化的发病机制。通过贴壁法用无血清培养基培养唾液腺上皮细胞,传三代后在IFN-γ及抗-CD40单抗分别孵育24h后收集细胞,采用流式细胞术、RT-PCR技术和免疫组化技术分析SS患者和健康对照组唾液腺上皮细胞CD40及ICAM-1的表达。流式细胞术、RT-PCR结果显示,在静止状态下,SS患者和健康对照组的唾液腺上皮细胞表达CD40及ICAM-1,经过IFN-γ及抗CD40单抗刺激后,CD40及ICAM-1的表达显著增强(P<0.01)。免疫组化结果显示,健康对照唾液腺基本不表达或低表达CD40和ICAM-1分子,而SS患者的唾液腺高表达CD40和ICAM-1分子。此外,经过IFN-γ及抗CD40单抗刺激后,SS组唾液腺上皮细胞的CD40及ICAM-1表达均显著高于对照组(P<0.01),提示SS唾液腺上皮细胞CD40及ICAM-1的高表达可能与其发病机制密切相关。
To investigate the difference of CD40, ICAM-1 / CD54 protein and gene expression in salivary gland epithelial cells between patients with Sjgrenren’s syndrome (SS) and healthy controls and to reveal the pathogenesis of abnormal immune activation of salivary gland epithelial cells in patients with SS. Salivary gland epithelial cells were cultured in serum-free medium by adherence method. After passage three generations, the cells were harvested respectively after incubation with IFN-γ and anti-CD40 mAb for 24 hours. Flow cytometry, RT-PCR and immunohistochemistry Expression of CD40 and ICAM-1 in salivary gland epithelial cells of SS patients and healthy controls. The results of flow cytometry and RT-PCR showed that the expression of CD40 and ICAM-1 in salivary gland epithelial cells of SS patients and healthy controls was significantly higher than that of control group after stimulation with IFN-γ and anti-CD40 mAbs The expression was significantly enhanced (P <0.01). The results of immunohistochemistry showed that the healthy control salivary glands did not express or down-regulate the expression of CD40 and ICAM-1, whereas the salivary glands of SS patients highly expressed CD40 and ICAM-1. In addition, the expressions of CD40 and ICAM-1 in salivary gland epithelial cells of SS group were significantly higher than those of control group (P <0.01) after stimulation with IFN-γ and anti-CD40 monoclonal antibody, suggesting that CD40 and ICAM-1 were high in SS salivary gland epithelial cells Expression may be closely related to its pathogenesis.