Basciano L K, Berenstein E H, Kmak L, Siraganian R P
J Biol Chem. 1986 Sep 5;261(25):11823-31.
Four monoclonal antibodies were produced that inhibit IgE binding to the high affinity IgE receptor (Fc epsilon R) on rat basophilic leukemia cells. The four monoclonal antibodies (mAb) fall into two groups. The first group was comprised of 3 antibodies (mAb BC4, mAb CD3, and mAb CA5) that reacted with the Fc epsilon R at epitopes close or identical to the IgE-binding site. With 125I-labeled antibodies there was reciprocal cross-inhibition between the antibodies and IgE. The antibodies activated both RBL-2H3 cells and normal rat mast cells for histamine release. The 3 antibodies immunoprecipitated the previously described alpha, beta, and gamma components of the receptor. The number of radiolabeled Fab fragments of 2 of these antibodies bound per cell was similar or equal to the number of IgE receptors. In contrast, the mAb BC4 Fab bound to 2.1 +/- 0.4 times the number of IgE receptor sites. Therefore, the portion of the Fc epsilon R exposed on the cell surface must have two identical epitopes and an axis of symmetry. These 3 monoclonal antibodies recognize different but closely related epitopes in the IgE-binding region of the Fc epsilon R. The fourth monoclonal antibody (mAb AA4) had different characteristics. In cross-inhibition studies, IgE and the other 3 monoclonals did not inhibit the binding of this 125I-labeled monoclonal antibody. The number of molecules of this antibody bound per cell was approximately 14-fold greater than the Fc epsilon R number. This monoclonal antibody caused the inhibition of histamine release and it appears to bind to several cell components.
制备了四种单克隆抗体,它们可抑制IgE与大鼠嗜碱性白血病细胞上的高亲和力IgE受体(FcεR)结合。这四种单克隆抗体(mAb)分为两组。第一组由3种抗体(mAb BC4、mAb CD3和mAb CA5)组成,它们在与IgE结合位点相近或相同的表位与FcεR发生反应。用125I标记抗体时,这些抗体与IgE之间存在相互交叉抑制。这些抗体可激活RBL-2H3细胞和正常大鼠肥大细胞释放组胺。这3种抗体免疫沉淀了先前描述的受体的α、β和γ组分。每细胞结合的这2种抗体的放射性标记Fab片段数量与IgE受体数量相似或相等。相比之下,mAb BC4 Fab结合的IgE受体位点数量是其2.1±0.4倍。因此,细胞表面暴露的FcεR部分必须有两个相同的表位和一个对称轴。这3种单克隆抗体识别FcεR的IgE结合区域中不同但密切相关的表位。第四种单克隆抗体(mAb AA4)具有不同的特性。在交叉抑制研究中,IgE和其他3种单克隆抗体不抑制这种125I标记的单克隆抗体的结合。每细胞结合的这种抗体分子数量比FcεR数量大约多14倍。这种单克隆抗体可抑制组胺释放,并且它似乎与几种细胞成分结合。