IDENTIFICATION OF THE C3BI RECEPTOR OF HUMAN-MONOCYTES AND MACROPHAGES BY USING MONOCLONAL-ANTIBODIES
IDENTIFICATION OF THE C3BI RECEPTOR OF HUMAN-MONOCYTES AND MACROPHAGES BY USING MONOCLONAL-ANTIBODIES
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DOI：
10.1073/pnas.80.18.5699
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发表时间：
1983-01-01
期刊：
影响因子：
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通讯作者：
SILVERSTEIN, SC
中科院分区：
文献类型：
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作者：
WRIGHT, SD;RAO, PE;SILVERSTEIN, SC
四种单克隆抗体IB 4、OKM 1、OKM 9和OKM 10均针对人单核细胞和巨噬细胞（M.vphi.）获得了使用两个标准来确定这些抗体的特异性。首先，培养物表面包被的抗受体抗体引起特异性下调的C3 bi受体活性的M. vphi。附着在这些基质上。其次，通过使用IB 4或OKM 1纯化的受体蛋白保留了选择性结合C3 bi包被的红细胞的能力。每种抗体识别C3 bi受体上的不同表位;它们不相互竞争结合单核细胞。当固定在固体支持物上时，每种抗体结合来自M. vphi的分子。可以同时结合其它单克隆抗C3 bi受体抗体之一的裂解物。OKM 10结合并掩蔽C3 bi受体的配体结合位点，而IB 4、OKM 1和OKM 9结合远离C3 bi结合位点的位点。所有4种抗体免疫沉淀来自125 I-表面标记的M. vphi的MW为185，000 [道尔顿]和105，000的多肽。IB 4还沉淀MW为185，000、153，000和105，000的多肽。显然，人M. vphi.是由两种多肽组成的复合物，MW 185，000和105，000。鉴定了与该复合物的4种不同抗原决定簇反应的单克隆抗体。抗体OKM 10识别的决定簇位于或靠近受体的配体结合位点。由抗体IB 4识别的决定簇被至少2种其它白细胞表面蛋白共享。
Four monoclonal antibodies, IB4, OKM1, OKM9 and OKM10, all directed against the C3bi [complement component 3b inhibitor] receptor of human monocytes and macrophages (M.vphi.) were obtained. Two criteria were used to determine the specificity of these antibodies. First, culture surfaces coated with the antireceptor antibodies caused specific down modulation of C3bi receptor activity on M.vphi. adherent to these substrates. Second, receptor protein purified by using IB4 or OKM1 retained the ability to bind selectively to C3bi-coated erythrocytes. Each of the antibodies recognizes a distinct epitope on the C3bi receptor; they do not compete with one another for binding to monocytes. When immobilized on a solid support, each of the antibodies binds a molecule from M.vphi. lysates that can simultaneously bind one of the other monoclonal anti-C3bi receptor antibodies. OKM10 binds and masks the ligand-binding site of the C3bi receptor, while IB4, OKM1 and OKM9 bind to sites remote from the C3bi binding site. All 4 antibodies immunoprecipitated polypeptides of MW 185,000 [daltons] and 105,000 from 125I-surface-labeled M.vphi.. IB4 also precipitates polypeptides of MW 185,000, 153,000 and 105,000. Evidently, the C3bi receptor of human M.vphi. is a complex composed of 2 polypeptides, MW 185,000 and 105,000. Monoclonal antibodies reacting with 4 distinct antigenic determinants of this complex were identified. The determinant recognized by antibody OKM10 is at or near the ligand-binding site of the receptor. The determinant recognized by antibody IB4 is shared by at least 2 other leukocyte surface proteins.