Olszowy M W, Leuchtmann P L, Veillette A, Shaw A S
Center for Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
J Immunol. 1995 Nov 1;155(9):4236-40.
The expression of the src-family kinases, p56lck and p59fyn, is critical for thymocyte development and TCR-mediated signal transduction, and may be important for signaling through other lymphoid receptors as well. Overexpression studies have demonstrated that the levels of p56lck and p59fyn expression can affect T cell development and signaling through the TCR. Therefore, it is likely that their exact expression levels play an important role in modulating signaling in thymocytes, mature T cells, and other lymphocytes. Here, we used quantitative immunoblotting to measure p56lck and p59fyn protein expression levels in thymocyte subsets, peripheral T cells, NK cells, and lymphoid cell lines. p59fyn expression levels were similar to p56lck in most cells that were examined demonstrating that p59fyn is abundantly expressed in T cells. In addition, we found that p56lck protein expression is equivalent in CD4 and CD8 double-negative, double-positive, and single-positive thymocytes. In contrast, p59fyn expression levels were significantly lower in double-positive thymocytes than in the other thymocyte subpopulations. Finally, we demonstrate that p56lck and p59fyn expression varies greatly in a number of cell lines used to study T cell activation and that IL-2 treatment can dynamically regulate p56lck and p59fyn expression in some cells.
src家族激酶p56lck和p59fyn的表达对胸腺细胞发育和TCR介导的信号转导至关重要,对通过其他淋巴受体的信号传导可能也很重要。过表达研究表明,p56lck和p59fyn的表达水平可影响T细胞发育及通过TCR的信号传导。因此,它们的确切表达水平可能在调节胸腺细胞、成熟T细胞及其他淋巴细胞的信号传导中发挥重要作用。在此,我们采用定量免疫印迹法来测定胸腺细胞亚群、外周T细胞、NK细胞及淋巴样细胞系中p56lck和p59fynyn的蛋白表达水平。在大多数检测的细胞中,p59fyn的表达水平与p56lck相似,表明p59fyn在T细胞中大量表达。此外,我们发现p56lck蛋白在CD4和CD8双阴性、双阳性及单阳性胸腺细胞中的表达相当。相比之下,双阳性胸腺细胞中p59fyn的表达水平显著低于其他胸腺细胞亚群。最后,我们证明在一些用于研究T细胞活化的细胞系中,p56lck和p59fyn的表达差异很大,并且白细胞介素-2处理可动态调节某些细胞中p56lck和p59fyn的表达。