Role of Crk-associated substrate lymphocyte type in the pathophysiology of rheumatoid arthritis in tax transgenic mice and in humans
Role of Crk-associated substrate lymphocyte type in the pathophysiology of rheumatoid arthritis in tax transgenic mice and in humans
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DOI：
10.1002/art.11047
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发表时间：
2003-07-01
影响因子：
--
通讯作者：
Morimoto, C
中科院分区：
文献类型：
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作者：
Miyake-Nishijima, R;Iwata, S;Morimoto, C
目标。目的探讨β1整合素下游信号分子Cas-L在类风湿关节炎(RA)发病机制中的作用。我们分析了人类T淋巴细胞病毒I型(HTLV-I)TAX转基因小鼠以及来自人类RA患者的样本。将来自TAX转基因小鼠的脾细胞培养在小鼠内皮细胞覆盖的Transwell插入物上，并计数通过内皮细胞单层的迁移细胞。用生化方法分析Cas-L的蛋白表达和酪氨酸磷酸化。用免疫组织化学方法检测Cas-L阳性细胞是否已渗入关节内。TAX转基因关节炎小鼠(ATG)的脾细胞迁移活性明显高于非关节炎TAX转基因小鼠(NTG)和同龄对照小鼠。与正常对照组和正常对照组相比，糖尿病模型组小鼠CaS-L蛋白表达及其酪氨酸磷酸化水平显著升高，并伴随着Fyn和Lck自身磷酸化水平的增强。免疫组织化学分析显示大量CaS-L阳性淋巴细胞迁移至患侧关节。此外，在人类类风湿关节炎中，Cas-L阳性淋巴细胞可渗透到炎性病变中。这些结果有力地提示CaS-L在RA的病理生理过程中起重要作用。
Objective. To investigate the role of Crk-associated substrate lymphocyte type (Cas-L), a downstream signaling molecule of beta1 integrins, in the pathophysiology of rheumatoid arthritis (RA).Methods. We analyzed human T lymphotropic virus type I (HTLV-I) tax transgenic mice as well as samples from human RA patients. Splenocytes from tax transgenic mice were cultured on mouse endothelial cell-covered Transwell inserts, and cells migrating through the endothelial monolayer were counted. Biochemical studies were performed to analyze the protein expression and tyrosine phosphorylation of Cas-L. Immunohistochemical analysis was performed to detect Cas-L-positive cells that had infiltrated into the joints.Results. Migratory activity of splenocytes from tax transgenic mice with arthritis (ATg) was much higher than that of tax transgenic mice without arthritis (NTg) and littermate control mice. The expression of Cas-L protein and its tyrosine phosphorylation were increased in ATg mice compared with NTg and control mice, and this was accompanied by enhanced autophosphorylation of Fyn and Lck. Immunohistochemical analysis demonstrated a large number of Cas-L-positive lymphocytes migrating into the affected joints. Furthermore, in human RA, Cas-L-positive lymphocytes were shown to infiltrate to the inflammatory lesions.Conclusion. These results strongly suggest that Cas-L plays an important role in the pathophysiology of RA.