Novel anti-cholesterol monoclonal immunoglobulin G antibodies as probes and potential modulators of membrane raft-dependent immune functions
Novel anti-cholesterol monoclonal immunoglobulin G antibodies as probes and potential modulators of membrane raft-dependent immune functions
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DOI：
10.1194/jlr.m600158-jlr200
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发表时间：
2007-01-01
影响因子：
6.5
通讯作者：
Laszlo, Gloria
中科院分区：
文献类型：
--
作者：
Biro, Adrienn;Cervenak, Laszlo;Laszlo, Gloria
L天然的抗胆固醇自身抗体存在于所有健康人的血清中，但其功能、产生和调节尚不清楚。在这里，我们通过用富含胆固醇的脂质体免疫小鼠，成功地制备了两种抗胆固醇的单抗(AchA)。新的AchA对胆固醇和一些结构上密切相关的3β-羟基甾醇具有特异性，它们可以与人的脂蛋白VLDL、LDL和HDL反应。它们通常以较低的亲和力结合到活的人类或小鼠淋巴细胞和单核巨噬细胞系上，细胞表面适度的木瓜酶消化大大增强了这种结合，去除了一些突出的细胞外蛋白结构域。与细胞结合的Achas与富含胆固醇的脂筏和小凹的标记在细胞表面以及细胞内与内质网和高尔基复合体的标记强烈地共存。这些数据表明，这些免疫球蛋白AchA可能作为活细胞中聚集的胆固醇(例如，不同的脂筏)的探针，因此也可能具有免疫调节潜力。
l Natural autoantibodies against cholesterol are present in the sera of all healthy individuals; their function, production, and regulation, however, are still unclear. Here, we managed to produce two monoclonal anti-cholesterol antibodies (ACHAs) by immunizing mice with cholesterol-rich liposomes. The new ACHAs were specific to cholesterol and to some structurally closely related 3 beta-hydroxyl sterols, and they reacted with human lipoproteins VLDL, LDL, and HDL. They bound, usually with low avidity, to live human or murine lymphocyte and monocyte-macrophage cell lines, which was enhanced substantially by a moderate papain digestion of the cell surface, removing some protruding extracellular protein domains. Cell-bound ACHAs strongly colocalized with markers of cholesterol-rich lipid rafts and caveolae at the cell surface and intracellularly with markers of the endoplasmic reticulum and Golgi complex. These data suggest that these IgG ACHAs may serve as probes of clustered cholesterol (e. g., different lipid rafts) in live cells and thus may also have immunomodulatory potential.