Haziot A, Rong G W, Bazil V, Silver J, Goyert S M
Division of Molecular Medicine, North Shore University Hospital/Cornell University Medical College, Manhasset, NY 11030.
J Immunol. 1994 Jun 15;152(12):5868-76.
CD14 functions as a cell surface receptor for LPS in the activation of monocytes/macrophages and neutrophils by endotoxin. To assess the utility of soluble forms of the CD14 receptor as a possible therapeutic for endotoxin shock, we have produced recombinant human soluble CD14 using a baculovirus expression system. We find that the recombinant protein is not only expressed on the surface of the insect cells as a glycosyl phosphatidylinositol (GPI)-anchored protein, but is also released into the culture medium as a soluble form that lacks the GPI anchor. Functional analyses of recombinant human soluble CD14 show that it binds specifically to LPS and can inhibit the LPS-induced release of TNF-alpha by macrophages and mononuclear cells as well as by cells in whole human blood when used at concentrations of approximately 70 micrograms/ml. Thus, soluble CD14 may be useful as an adjunct in the treatment of endotoxin shock.
在单核细胞/巨噬细胞和中性粒细胞被内毒素激活的过程中,CD14作为脂多糖的细胞表面受体发挥作用。为了评估可溶性形式的CD14受体作为内毒素休克可能治疗方法的效用,我们使用杆状病毒表达系统制备了重组人可溶性CD14。我们发现,重组蛋白不仅作为糖基磷脂酰肌醇(GPI)锚定蛋白表达在昆虫细胞表面,还以缺乏GPI锚的可溶性形式释放到培养基中。重组人可溶性CD14的功能分析表明,当以约70微克/毫升的浓度使用时,它能特异性结合脂多糖,并可抑制巨噬细胞、单核细胞以及全血中的细胞由脂多糖诱导的肿瘤坏死因子-α释放。因此,可溶性CD14可能作为内毒素休克治疗的辅助药物有用。