English A, Kosoy R, Pawlinski R, Bamezai A
Department of Cellular Biology University of Georgia, Athens 30602, USA.
J Immunol. 2000 Oct 1;165(7):3763-71. doi: 10.4049/jimmunol.165.7.3763.
The Ly-6 locus encodes several cell surface proteins of 10-12 kDa. Some members of this multigene family may function in cell signaling and/or cell adhesion processes. T lymphocytes overexpressing Ly-6A.2 (one member of the Ly-6 gene family) protein homotypically aggregate when cultured in vitro. Further analysis of this homotypic aggregation suggests that Ly-6A.2 participates in cell-cell adhesion. These observations indicated the presence of a Ly-6 ligand(s) on the surface of lymphoid cells. In this study we report generation of a hamster mAb, 9AB2, that blocks Ly-6A.2-dependent cell-cell adhesion. The 9AB2 Ab recognizes a 66-kDa glycoprotein with unique tissue expression. The 9AB2 mAb does not bind Ly-6A.2, but coimmunoprecipitates Ly-6A.2 molecule. Moreover, 9AB2 Ag-expressing thymocytes specifically bind to Chinese hamster ovary cells overexpressing Ly-6A.2 protein, and this binding is specifically blocked by 9AB2 and anti-Ly-6A.2 Abs. These results suggest that the 66-kDa protein recognized by 9AB2 mAb is the putative ligand for Ly-6A.2.
Ly-6基因座编码几种10 - 12 kDa的细胞表面蛋白。这个多基因家族的一些成员可能在细胞信号传导和/或细胞黏附过程中发挥作用。过表达Ly-6A.2(Ly-6基因家族的一个成员)蛋白的T淋巴细胞在体外培养时会发生同型聚集。对这种同型聚集的进一步分析表明,Ly-6A.2参与细胞间黏附。这些观察结果表明淋巴细胞表面存在Ly-6配体。在本研究中,我们报告了一种仓鼠单克隆抗体9AB2的产生,它能阻断Ly-6A.2依赖的细胞间黏附。9AB2抗体识别一种具有独特组织表达的66 kDa糖蛋白。9AB2单克隆抗体不与Ly-6A.2结合,但能共免疫沉淀Ly-6A.2分子。此外,表达9AB2抗原的胸腺细胞能特异性结合过表达Ly-6A.2蛋白的中国仓鼠卵巢细胞,并且这种结合被9AB2和抗Ly-6A.2抗体特异性阻断。这些结果表明9AB2单克隆抗体识别的66 kDa蛋白是Ly-6A.2的假定配体。