Functional Analyses of Patient-derived IgG Monoclonal Anticardiolipin Antibodies Using In Vivo Thrombosis and In Vivo Microcirculation Models
Functional Analyses of Patient-derived IgG Monoclonal Anticardiolipin Antibodies Using In Vivo Thrombosis and In Vivo Microcirculation Models
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DOI：
10.1055/s-0037-1614033
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发表时间：
2000-09
影响因子：
6.7
通讯作者：
S. Pierangeli;Xiaowei Liu;R. Espinola;Tsawei Olee;Min Zhu;N. Harris;Pojen P. Chen
中科院分区：
文献类型：
--
作者：
S. Pierangeli;Xiaowei Liu;R. Espinola;Tsawei Olee;Min Zhu;N. Harris;Pojen P. Chen
抗磷脂抗体（aPL）与抗磷脂综合征（APS）患者的血栓形成和妊娠丢失有关，并可增强小鼠体内血栓形成，但aPL导致血栓形成的机制尚未完全了解。已经提出aPL可能影响内皮细胞（EC）功能和/或诱导其活化，将其抗凝剂表面转化为促凝血剂，从而诱发血栓形成。已经提出aPL可能影响EC细胞功能和/或诱导其活化，将其抗凝剂表面转化为促凝血剂，从而诱发血栓形成。本研究旨在验证一些IgG抗心磷脂（IgG aCL）在小鼠体内具有血栓形成特性，通过激活内皮发挥其作用的假设。我们研究了7个患者源性单克隆ACL的血栓形成的特性在体内捏诱导的血栓形成模型，并通过分析在体内白细胞粘附内皮微循环暴露小鼠提睾肌和体外粘附分子表达培养EC激活EC的功能活动。还测试了单克隆aCL与EC的结合。除了先前确定的血栓形成IS 2外，发现5个新的IgG单克隆aCL（来自2名患者）中有4个具有血栓形成性。在这五种血栓形成性ACL中，有三种引起了体内白细胞粘附于微循环中的EC，与H2对照人单克隆IgG诱导的粘附相比，并增强了粘附分子（特别是VCAM-1）在培养EC上的表达。这些数据表明，约2/3的患者源性IgG单克隆aCL具有血栓形成性，并提示某些血栓形成性IgG aCL通过激活EC发挥其作用。缩略语名称：aCL，抗心磷脂抗体; aPL，抗磷脂抗体; APS，抗磷脂综合征; CL，心磷脂; dRVVT，稀释罗素蝰蛇毒时间; EC，内皮细胞; ELISA，酶联免疫吸附测定; GP，糖蛋白; h，小时; HUVEC，人脐静脉内皮细胞; ICAM，细胞间粘附分子; KCT，高岭土凝血时间; LA，狼疮抗凝剂; PBS，磷酸盐缓冲盐水; PL，磷脂; sq，正方形; TBS，Tris缓冲盐水; VCAM，血管细胞粘附分子; PMBC，外周血单核细胞
Summary Antiphospholipid antibodies (aPL) have been associated with thrombosis and pregnancy losses in patients diagnosed with antiphospholipid syndrome (APS) and enhance thrombus formation in vivo in mice, but the mechanism of thrombosis by aPL is not completely understood. It has been proposed that aPL may affect endothelial cell (EC) function and/or induce their activation, transforming their anticoagulant surface into procoagulant, thus predisposing to thrombosis. It has been proposed that aPL may affect EC cell function and/or induce their activation, transforming their anticoagulant surface into procoagulant, thus predisposing to thrombosis. This study proposes to test the hypotheses that some IgG anticardiolipins (IgG aCL) with thrombogenic properties in mice, exert their effects through activation of endothelium. We studied seven patient-derived monoclonal aCL for their thrombogenic properties in an in vivo pinch-induced thrombosis model, and their functional activities in activating EC by analyzing in vivo leukocyte adhesion to endothelium in microcirculation in venules in exposed murine cremaster muscle and in vitro adhesion molecule expression in cultured EC. The binding of the monoclonal aCL to EC was also tested. In addition to the previous identified thrombogenic IS2, four of the five new more IgG monoclonal aCL (from two patients) were found to be thrombogenic. Of these five thrombogenic aCL, three caused more in vivo leukocyte adhesion to EC in microcirculation, as compared to that induced by the H2 control human monoclonal IgG, and enhanced expression of adhesion molecules (particularly VCAM-1) on cultured EC. These data show that about 2/3 patientderived IgG monoclonal aCL are thrombogenic and suggest that some thrombogenic IgG aCL exert their effects through activating EC. Abbreviations: aCL, anticardiolipin antibodies; aPL, antiphospholipid antibodies; APS, antiphospholipid syndrome; CL, cardiolipin; dRVVT, dilute Russell’s viper venom time; EC, endothelial cells; ELISA, enzyme-linked immunosorbent assay; GP, glycoprotein; h, hour; HUVEC, human umbilicalvein endothelial cells; ICAM, intercellular adhesion molecule; KCT, kaolin clotting time; LA, lupus anticoagulant; PBS, phosphate-buffered saline; PL, phospholipid; sq, square; TBS, Tris-buffered saline; VCAM, vascular cell adhesion molecule; PMBC, peripheral mononuclear blood cells