Use of the monoclonal antibody 12F1 to characterize the differentiation antigen VLA-2.
Use of the monoclonal antibody 12F1 to characterize the differentiation antigen VLA-2.
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DOI：
10.4049/jimmunol.138.1.226
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发表时间：
1987-01
影响因子：
4.4
通讯作者：
K. Pischel;M. Hemler;C. Huang;H. Bluestein;V. Woods
中科院分区：
文献类型：
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作者：
K. Pischel;M. Hemler;C. Huang;H. Bluestein;V. Woods
已经产生了单克隆抗体12 F1，其特异性免疫沉淀来自血小板和长期活化的T细胞以及来自成纤维细胞和神经母细胞瘤细胞系的人细胞表面结构VLA-2。交联研究表明，VLA-2结构以165，000 Mr重链（α 2）与130，000 Mr轻链（β）以1：1非共价缔合的形式存在于细胞表面。与所有VLA结构共有的β亚基反应的单克隆抗体A-1A 5能够完全预清除VLA-2，表明所有α 2亚基与VLA β链相关。12 F1对VLA-2的特异性允许对该α 2 β结构进行独立的免疫沉淀和流式细胞术分析，该α 2 β结构与可能存在的任何其他VLA结构（如VLA-1或游离β亚基）分开。亚基解离研究用于证明12 F1识别VLA-2上α 2链上的表位，这与12 F1对VLA蛋白中单独的VLA-2的特异性一致。活化的T细胞的分析表明，VLA-2，像VLA-1，是另一种“非常晚”出现的T细胞活化抗原，其从第7天开始与VLA-1同时出现，并在2周内增加。VLA-2在许多与VLA-1相同的细胞（失活的T细胞、T细胞白血病细胞、成纤维细胞、SK-N-SH神经母细胞瘤细胞）上发现，但是VLA-1和VLA-2可以独立地表达，因为VLA-2也存在于VLA-1阴性细胞如HSB和血小板上，并且VLA-1存在于VLA-2阴性C8215细胞上。
A monoclonal antibody, 12F1, has been produced that specifically immunoprecipitates the human cell surface structure VLA-2 from platelets and long-term activated T cells, as well as from fibroblast and neuroblastoma cell lines. Cross-linking studies indicate that the VLA-2 structure exists on the cell surface as a 165,000 Mr heavy chain (alpha 2) in noncovalent 1:1 association with a 130,000 Mr light chain (beta). The monoclonal antibody A-1A5, which reacts with the beta subunit common to all VLA structures, was able to completely preclear VLA-2, indicating that all of the alpha 2 subunit was associated with VLA beta-chain. The specificity of 12F1 for VLA-2 allowed independent immunoprecipitation and flow cytometry analysis of this alpha 2 beta structure separate from any other VLA structures that may have been present such as VLA-1 or free beta-subunit. Subunit dissociation studies were used to demonstrate that 12F1 recognizes an epitope on the alpha 2 chain on VLA-2, which is consistent with the 12F1 specificity for VLA-2 alone among the VLA proteins. Analysis of activated T cells indicated that VLA-2, like VLA-1, is another "very late" appearing T cell activation antigen that arises concurrently with VLA-1 starting at day 7 and increasing through 2 wk. VLA-2 was found on many of the same cells as VLA-1 (inactivated T cells, T cell leukemia cells, fibroblasts, SK-N-SH neuroblastoma cells), but VLA-1 and VLA-2 can be expressed independently, because VLA-2 was also present on VLA-1-negative cells such as HSB and platelets, and VLA-1 was present on VLA-2-negative C8215 cells.