Grégoire C, Rebaï N, Schweisguth F, Necker A, Mazza G, Auphan N, Millward A, Schmitt-Verhulst A M, Malissen B
Centre d'Immunologie, Institut National de la Santé et de la Recherche Médicale, Marseille, France.
Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):8077-81. doi: 10.1073/pnas.88.18.8077.
We have produced a soluble form of a mouse alpha beta T-cell antigen receptor (TCR) by shuffling its variable (V) and constant (C) domains to the C region of an immunoglobulin kappa light chain. These chimeric molecules composed of V alpha C alpha C kappa and V beta C beta C kappa chains were efficiently secreted (up to 1 micrograms/ml) by transfected myeloma cells as noncovalent heterodimers of about 95-kDa molecular mass. In the absence of direct binding measurement, we have refined the epitopic analysis of the soluble V alpha C alpha C kappa-V beta C beta C kappa dimers and shown that they react with an anti-clonotypic antibody and two antibodies directed to the C domain of the TCR alpha and beta chains. Conversely, we have raised three distinct monoclonal antibodies against the soluble TCR heterodimers and shown that they recognize surface-expressed TCRs. Two of these antibodies were found to react specifically with the products of the V alpha 2 (V delta 8) and V beta 2 gene segments, respectively. When considered together, these data suggest that these soluble TCR molecules are folded in a conformation indistinguishable from that which they assume at the cell surface.
我们通过将小鼠αβ T细胞抗原受体（TCR）的可变（V）结构域和恒定（C）结构域与免疫球蛋白κ轻链的C区域进行重排，制备出了一种可溶性形式的TCR。这些由VαCαCκ和VβCβCκ链组成的嵌合分子作为约95 kDa分子量的非共价异二聚体，由转染的骨髓瘤细胞高效分泌（高达1微克/毫升）。在没有直接结合测量的情况下，我们对可溶性VαCαCκ-VβCβCκ二聚体进行了表位分析的优化，并表明它们能与抗独特型抗体以及两种针对TCRα和β链C结构域的抗体发生反应。相反，我们制备了三种针对可溶性TCR异二聚体的不同单克隆抗体，并表明它们能识别表面表达的TCR。发现其中两种抗体分别与Vα2（Vδ8）和Vβ2基因片段的产物特异性反应。综合考虑这些数据表明，这些可溶性TCR分子折叠成的构象与它们在细胞表面所呈现的构象无法区分。
