Basi G S, Riggs M B, Nash K, Singer R
Protein Design Labs, Inc., Mountain View, CA 94043.
J Immunol Methods. 1992 Nov 5;155(2):175-91. doi: 10.1016/0022-1759(92)90284-z.
The T cell receptor (TcR) is an integral membrane protein occurring as a disulfide linked heterodimer, non-covalently associated with CD3 on the surface of T lymphocytes. Antibodies to the TcR have been shown to be effective for treating autoimmune disorders in animals. We describe here a method for producing antibodies to cell surface determinants of the human TcR, using a soluble form of the receptor as antigen. Soluble V alpha 1.2, V beta 8.1, V beta 11 TcR chains are expressed from a construct in which the extracellular domains of the TcR are fused to the mouse gamma 2a heavy chain constant region lacking the CH1 domain. These chimeric molecules contain both immunoglobulin and TcR determinants, as revealed by antibody probes. Amino-terminal sequence analysis of a chimeric V beta 8.1 molecule indicates that the TcR leader peptide is correctly processed from the soluble form. Antibodies raised against the soluble human V beta 8.1 molecule recognize the native determinants on Jurkat cells, and on natural T cells derived from resting human peripheral blood lymphocytes. Epitope mapping studies using competitive binding assays suggest that the anti-V beta 8 antibodies produced using soluble antigen recognize multiple overlapping determinants on the cell surface form of the TcR.
T细胞受体（TcR）是一种整合膜蛋白，以二硫键连接的异二聚体形式存在，与T淋巴细胞表面的CD3非共价结合。已证明针对TcR的抗体对治疗动物自身免疫性疾病有效。我们在此描述一种方法，使用受体的可溶性形式作为抗原，生产针对人TcR细胞表面决定簇的抗体。可溶性Vα1.2、Vβ8.1、Vβ11 TcR链从一种构建体表达，其中TcR的胞外结构域与缺少CH1结构域的小鼠γ2a重链恒定区融合。如抗体探针所示，这些嵌合分子同时含有免疫球蛋白和TcR决定簇。对一种嵌合Vβ8.1分子的氨基末端序列分析表明，TcR前导肽从可溶性形式中正确加工。针对可溶性人Vβ8.1分子产生的抗体识别Jurkat细胞以及源自静息人外周血淋巴细胞的天然T细胞上的天然决定簇。使用竞争结合试验的表位作图研究表明，使用可溶性抗原产生的抗Vβ8抗体识别TcR细胞表面形式上的多个重叠决定簇。
