O'Grady J H, Looney R J, Anderson C L
J Immunol. 1986 Oct 1;137(7):2307-10.
The valence for ligand of the 72 kD high-affinity IgG FcR present on human mononuclear phagocytes was evaluated. Lysates of U937 cells whose high-affinity FcR had been saturated with equivalent quantities of 125I-IgG1 kappa and unlabeled IgG1 lambda or with 125I-IgG1 lambda and unlabeled IgG1 kappa were incubated with Sepharose-anti-kappa. Eighty-nine percent of the applied 125I-IgG1 kappa was bound, whereas 0.35% of the applied 125I-IgG1 lambda bound (mean of two experiments), indicating that if the receptors are occupied with ligand, the receptors bind only one ligand molecule at a time. Two experiments were performed to show that the receptors were ligand-occupied. First, a monoclonal antibody directed against the 72 kD FcR (FcRmab32) was added to lysates of U937 cells saturated with equal quantities of 125I-IgG1 lambda and IgG1 kappa. This anti-FcR antibody caused a dose-dependent sevenfold increase in the amount of 125I-IgG1 lambda bound to the anti-kappa immunoadsorbent (presumably by cross-linking receptors bearing 125I-IgG1 lambda with receptors bearing IgG1 kappa), whereas monoclonal antibodies (MMA and IV3) directed against two other determinants on U937 caused no such increase. In the second experiment, Sepharose-FcRmab32 adsorbed 60% of the 125I-IgG1 kappa and 46% of the 125I-IgG1 lambda applied in a U937 lysate (bearing high-affinity FcR), whereas only 3% of 125I-IgG1 kappa and 6% of 125I-IgG1 lambda applied in a K562 lysate (bearing no high-affinity FcR) were adsorbed. We interpret these data to indicate that in detergent solution the valency of the high-affinity FcR on U937 cells is one.
对人单核吞噬细胞上存在的72kD高亲和力IgG FcR的配体价进行了评估。将U937细胞的裂解物与琼脂糖-抗κ一起孵育,这些细胞的高亲和力FcR已被等量的125I-IgG1κ和未标记的IgG1λ或125I-IgG1λ和未标记的IgG1κ饱和。所施加的125I-IgG1κ中有89%被结合,而所施加的125I-IgG1λ中有0.35%被结合(两个实验的平均值),这表明如果受体被配体占据,受体一次仅结合一个配体分子。进行了两个实验以表明受体被配体占据。首先,将针对72kD FcR的单克隆抗体(FcRmab32)添加到用等量的125I-IgG1λ和IgG1κ饱和的U937细胞裂解物中。这种抗FcR抗体导致与抗κ免疫吸附剂结合的125I-IgG1λ量呈剂量依赖性增加7倍(可能是通过将携带125I-IgG1λ的受体与携带IgG1κ的受体交联),而针对U937上其他两个决定簇的单克隆抗体(MMA和IV3)则没有这种增加。在第二个实验中,琼脂糖-FcRmab32吸附了U937裂解物(携带高亲和力FcR)中所施加的125I-IgG1κ的60%和125I-IgG1λ的46%,而在K562裂解物(不携带高亲和力FcR)中所施加的125I-IgG1κ只有3%和125I-IgG1λ只有6%被吸附。我们解释这些数据表明在去污剂溶液中U937细胞上高亲和力FcR的价为一。