Murata Yoji, Saito Yasuyuki, Kaneko Tetsuya, Kotani Takenori, Kaneko Yoriaki, Ohnishi Hiroshi, Matozaki Takashi
Division of Molecular and Cellular Signaling, Department of Biochemistry and Molecular Biology, Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho, Chuo-ku, Kobe 650-0017, Japan.
Laboratory of Biosignal Sciences, Institute for Molecular and Cellular Regulation, Gunma University, 3-39-15 Showa-Machi, Maebashi, Gunma 371-8512, Japan; Department of Medicine and Clinical Science, Gunma University Graduate School of Medicine, 3-39-22 Showa-Machi, Maebashi, Gunma 371-8511, Japan.
Methods. 2014 Jan 15;65(2):254-9. doi: 10.1016/j.ymeth.2013.09.016. Epub 2013 Oct 1.
Signal regulatory protein α (SIRPα), also known as SHPS-1/SIRPA, is an immunoglobulin superfamily protein that binds to the protein tyrosine phosphatases Shp1 and Shp2 through its cytoplasmic region and is predominantly expressed in dendritic cells and macrophages. CD47, a widely expressed transmembrane protein, is a ligand for SIRPα, with the two proteins constituting a cell-cell communication system. It was previously demonstrated that the CD47-SIRPα signaling pathway is important for prevention of clearance by splenic macrophages of red blood cells or platelets from the bloodstream. In addition, this signaling pathway is also implicated in homeostatic regulation of dendritic cells and development of autoimmunity. Here we describe the detailed protocols for methods that were used in our recent studies to study the role of the CD47-SIRPα signaling pathway in autoimmunity. We also demonstrate that hematopoietic SIRPα as well as nonhematopoietic CD47 are important for development of experimental autoimmune encephalomyelitis. Thus, we here strengthen the importance of experimental animal models as well as other methods for the study of molecular pathogenesis of autoimmunity.
信号调节蛋白α（SIRPα），也称为SHPS-1/SIRPA，是一种免疫球蛋白超家族蛋白，通过其胞质区域与蛋白酪氨酸磷酸酶Shp1和Shp2结合，主要表达于树突状细胞和巨噬细胞中。CD47是一种广泛表达的跨膜蛋白，是SIRPα的配体，这两种蛋白构成了一个细胞间通讯系统。先前已证明，CD47-SIRPα信号通路对于防止脾脏巨噬细胞清除血液中的红细胞或血小板很重要。此外，该信号通路还参与树突状细胞的稳态调节和自身免疫的发展。在此，我们描述了我们最近研究中用于研究CD47-SIRPα信号通路在自身免疫中作用的方法的详细方案。我们还证明，造血SIRPα以及非造血CD47对于实验性自身免疫性脑脊髓炎的发展很重要。因此，我们在此强化了实验动物模型以及其他方法对于研究自身免疫分子发病机制的重要性。
