Punt J A, Roberts J L, Kearse K P, Singer A
Experimental Immunology Branch, National Cancer Institute, Bethesda, Maryland 20892.
J Exp Med. 1994 Aug 1;180(2):587-93. doi: 10.1084/jem.180.2.587.
The stoichiometry of the subunits that comprise the T cell antigen receptor (TCR) complex is not completely known. In particular, it is uncertain whether TCR alpha and TCR beta proteins are present in the TCR complex as one or multiple heterodimeric pairs. In this study we have used mice transgenic for two different TCR alpha and two different TCR beta proteins to determine the number of TCR alpha and TCR beta chains in a single TCR complex. Individual thymocytes and splenic T cells from double TCR transgenic mice simultaneously expressed all four transgenic TCR proteins on their surfaces. Because the individual TCR alpha and individual TCR beta proteins were biochemically distinguishable, we were able to examine association among the transgenic TCR products. We found that each TCR alpha chain paired with each TCR beta chain, but that each TCR complex contained only one TCR alpha and one TCR beta protein. Furthermore, quantitative immunofluorescence revealed that T cells expressed twice as many CD3 epsilon as TCR beta proteins. These findings demonstrate that there are precisely one TCR alpha, one TCR beta, and two CD3 epsilon chains in each TCR/CD3 complex expressed on the surfaces of both thymocytes and mature T cells.
构成T细胞抗原受体(TCR)复合物的亚基化学计量尚未完全明确。特别是,TCRα和TCRβ蛋白在TCR复合物中是以一个还是多个异源二聚体对的形式存在尚不确定。在本研究中,我们使用了转染了两种不同TCRα和两种不同TCRβ蛋白的转基因小鼠,以确定单个TCR复合物中TCRα和TCRβ链的数量。来自双TCR转基因小鼠的单个胸腺细胞和脾T细胞在其表面同时表达所有四种转基因TCR蛋白。由于单个TCRα和单个TCRβ蛋白在生化上是可区分的,我们能够检测转基因TCR产物之间的关联。我们发现每个TCRα链与每个TCRβ链配对,但每个TCR复合物仅包含一个TCRα和一个TCRβ蛋白。此外,定量免疫荧光显示T细胞表达的CD3ε是TCRβ蛋白的两倍。这些发现表明,在胸腺细胞和成熟T细胞表面表达的每个TCR/CD3复合物中,精确地存在一个TCRα、一个TCRβ和两条CD3ε链。