Generation and characterization of a novel adhesion function blocking monoclonal antibody recognizing both rat and mouse E-selectin.
Generation and characterization of a novel adhesion function blocking monoclonal antibody recognizing both rat and mouse E-selectin.
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识别大鼠和小鼠 E-选择素的新型粘附功能阻断单克隆抗体的生成和表征。
DOI：
10.1089/hyb.1997.16.355
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
Issekutz,AC
中科院分区：
文献类型：
--
作者：
Walter,UM;Ayer,LM;Manning,AM;Frenette,PS;Wagner,DD;Hynes,RO;Wolitzky,BA;Issekutz,AC
循环中的白细胞重新聚集到炎症部位的先决条件是与血管内皮细胞的黏附。选择素通过调节白细胞在内皮上的“滚动”，在这一多步骤过程的启动中发挥着重要作用。使用功能阻断单抗(MAb)研究选择素依赖的细胞相互作用，有助于深入了解白细胞迁移至炎症的机制。到目前为止，大鼠炎症模型的研究大多依赖于大鼠E-选择素的交叉反应抗体或多克隆抗体。在E-选择素基因敲除小鼠中，我们的目的是通过免疫大鼠E-选择素基因敲除的中国仓鼠卵巢细胞(RESEC)，产生一种阻断大鼠E-选择素单抗的黏附功能。经酶联免疫吸附试验证实，该κ能与RESec反应，但不能与未转基因的中国仓鼠卵巢细胞反应，也不能与重组小鼠E-选择素蛋白反应。这种单抗被命名为RME-1。与人脐静脉内皮细胞表达的大鼠L-选择素、大鼠P-选择素、E-选择素无交叉反应。在静态条件下，单抗RME-1可完全抑制HL-60髓系细胞与固定化小鼠E-选择素的黏附，在旋转条件下可完全阻断大鼠多形核白细胞与重组小鼠E-选择素的黏附。因此，这种新型抗体识别啮齿动物E-选择素的功能相关表位。
The prerequisite for the recruitment of circulating leukocytes to sites of inflammation is adhesion to vascular endothelial cells. Selectins play a significant role in the initiation of this multistep process by mediating "rolling" of the leukocytes on the endothelium. Investigation of selectin-dependent cell interactions using function blocking monoclonal antibodies (MAb) provides insights into the mechanisms involved in leukocyte migration into inflammation. Until now most studies in inflammation models in rats have relied on cross-reactive or polyclonal antibodies against rat E-selectin. In an E-selectin knockout mouse, we aimed to generate an adhesion function blocking MAb to rat E-selectin by immunization with rat E-selectin transfected Chinese hamster ovary cells (RESEC). An IgG1κ was identified that reacts with RESEC but not with untransfected Chinese hamster ovary cells, as well as with recombinant mouse E-selectin protein as assessed by ELISA. This MAb is designated RME-1. It does not cross-react with rat L-selectin or rat P-selectin or E-selectin expressed on human umbilical vein endothelium. Adhesion of the HL-60 myeloid cells to immobilized mouse E-selectin was completely inhibited by MAb RME-1 under static conditions and adhesion of rat polymorphonuclear leukocytes to recombinant mouse E-selectin was blocked under rotation conditions. This novel antibody thus recognizes a function-related epitope on rodent E-selectin.
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