Z Jiao1, W Wang, M Guo, T Zhang, L Chen, Y Wang, H You, J Li. 1. Department of Laboratory Medicine, Affiliated Hospital of Jiangsu University, Zhenjiang, PR China. jiaozhijun@ujs.edu.cn
Abstract
OBJECTIVE: Expression of Notch homologues in local tissue inflammation in rheumatoid arthritis (RA) and cultured synoviocytes has been reported, but the expression profile of Notch-related molecules in peripheral lymphocytes in RA remains unclear. In this study, we measured the expression of Notch receptors and downstream molecules in peripheral lymphocytes from RA patients. METHODS: Expression of Notch receptors in peripheral lymphocytes of RA patients was assessed by both flow cytometry and real-time polymerase chain reaction (PCR). Expression of the representative Notch target gene HES-1 and the regulatory gene NUMB in purified T helper cells from RA patients was determined by real-time PCR, and expression of Notch intracellular domain (ICD) was determined by immunoblot analysis. RESULTS: There was an increased expression of Notch 2, Notch 3, and Notch 4 in T helper cells from active RA patients, among which increased expression of Notch 3 was mainly by activated T cells. Notably, expression of Notch 3 in T cells decreased in inactive RA patients and the level was similar to that of healthy controls (HC). Notch receptors were rarely observed on B cells and no difference in expression was found between RA patients and HC. T helper cells from RA patients exhibited increased expression of the target gene HES-1 but decreased expression of the negative modulation gene NUMB of Notch signalling. There was also an increased nuclear translocation of Notch-ICD in T helper cells from active RA disease. CONCLUSION: The present study demonstrated that T helper cells from RA patients display a significantly altered expression profile of Notch receptors and enhanced activation of Notch signalling compared with HC.
OBJECTIVE: Expression of Notch homologues in local tissue inflammation in rheumatoid arthritis (RA) and cultured synoviocytes has been reported, but the expression profile of Notch-related molecules in peripheral lymphocytes in RA remains unclear. In this study, we measured the expression of Notch receptors and downstream molecules in peripheral lymphocytes from RApatients. METHODS: Expression of Notch receptors in peripheral lymphocytes of RApatients was assessed by both flow cytometry and real-time polymerase chain reaction (PCR). Expression of the representative Notch target gene HES-1 and the regulatory gene NUMB in purified T helper cells from RApatients was determined by real-time PCR, and expression of Notch intracellular domain (ICD) was determined by immunoblot analysis. RESULTS: There was an increased expression of Notch 2, Notch 3, and Notch 4 in T helper cells from active RApatients, among which increased expression of Notch 3 was mainly by activated T cells. Notably, expression of Notch 3 in T cells decreased in inactive RApatients and the level was similar to that of healthy controls (HC). Notch receptors were rarely observed on B cells and no difference in expression was found between RApatients and HC. T helper cells from RApatients exhibited increased expression of the target gene HES-1 but decreased expression of the negative modulation gene NUMB of Notch signalling. There was also an increased nuclear translocation of Notch-ICD in T helper cells from active RA disease. CONCLUSION: The present study demonstrated that T helper cells from RApatients display a significantly altered expression profile of Notch receptors and enhanced activation of Notch signalling compared with HC.