Neidhart M, Zaucke F, von Knoch R, Jüngel A, Michel B A, Gay R E, Gay S
Centre for Experimental Rheumatology, University Hospital, Gloriastrasse 25, CH-8091 Zurich, Switzerland.
Ann Rheum Dis. 2005 Mar;64(3):419-24. doi: 10.1136/ard.2004.023135. Epub 2004 Sep 2.
Galectin-3 is expressed in the synovial tissue of patients with rheumatoid arthritis (RA), particularly at sites of joint destruction.
To explore the possibilities that galectin-3 is induced either by proinflammatory cytokines or by adhesion to cartilage components.
Cell culture plates were coated with fibronectin, collagens I-VI, or cartilage oligomeric matrix protein (COMP), and the suspended cells were then added. The medium was changed after 1 hour at 37 degrees C. Adherent cells were further incubated for 18 hours in the presence or absence of tumour necrosis factor alpha (TNF alpha) or interleukin 1 beta. Cells were pretreated with murine IgG1, anti-CD29, -CD51, -CD61 (integrins), or -CD3 monoclonal antibodies and transferred to culture plates coated with COMP. Adherent cells were counted by light microscopy. The expression of intracellular galectin-3, or cell surface CD29, CD51, and CD61 was determined by flow cytometry before and after adhesion.
Four times more RA synovial fibroblasts (SF) than osteoarthritis SF adhered to COMP. RA SF presented more cell surface integrins, and monoclonal antibodies against CD51 inhibited the adhesion to COMP by 80%. TNF alpha reduced the expression of CD61 and the adhesion to COMP, but did not reverse the adhesion once it had taken place. The adhesion of RA SF to COMP was found to increase the intracellular level of galectin-3. In contrast, intracellular galectin-3 decreased after exposure to TNF alpha.
The increase of galectin-3 occurs after adhesion to COMP, and the alpha V beta 3 receptor (CD51/CD61) has a pivotal role in this process.
半乳糖凝集素-3在类风湿关节炎（RA）患者的滑膜组织中表达，尤其在关节破坏部位。
探讨促炎细胞因子或与软骨成分黏附是否可诱导半乳糖凝集素-3表达。
在细胞培养板上包被纤连蛋白、I-VI型胶原蛋白或软骨寡聚基质蛋白（COMP），然后加入悬浮细胞。37℃孵育1小时后更换培养基。贴壁细胞在有或无肿瘤坏死因子α（TNFα）或白细胞介素1β的情况下进一步孵育18小时。细胞用鼠IgG1、抗CD29、-CD51、-CD61（整合素）或-CD3单克隆抗体预处理，然后转移至包被有COMP的培养板上。通过光学显微镜计数贴壁细胞。在黏附前后，通过流式细胞术测定细胞内半乳糖凝集素-3或细胞表面CD29、CD51和CD61的表达。
RA滑膜成纤维细胞（SF）黏附于COMP的数量是骨关节炎SF的4倍。RA SF呈现更多的细胞表面整合素，抗CD51单克隆抗体可使黏附于COMP的能力降低80%。TNFα降低CD61的表达及对COMP的黏附，但在黏附发生后并不能逆转黏附。发现RA SF与COMP的黏附可增加细胞内半乳糖凝集素-3的水平。相反，暴露于TNFα后细胞内半乳糖凝集素-3减少。
黏附于COMP后半乳糖凝集素-3增加，αVβ3受体（CD51/CD61）在此过程中起关键作用。
