Ziegler-Heitbrock H W, Appl B, Käfferlein E, Löffler T, Jahn-Henninger H, Gutensohn W, Nores J R, McCullough K, Passlick B, Labeta M O
Institute for Immunology, Klinikum Innenstadt, Munich, Germany.
Scand J Immunol. 1994 Nov;40(5):509-14. doi: 10.1111/j.1365-3083.1994.tb03497.x.
Several monoclonal antibodies directed against the human CD14 antigen have been established. We now report that the antibody My4, but not LeuM3, reacts with porcine monocytes. Among porcine peripheral blood mononuclear cells (PBMC), 14.6% of the cells stain with the CD14 antibody My4, which is similar to the percentage obtained with the antiporcine monocyte antibody 74-22-15. Two-colour immunofluorescence reveals that My4 and 74-22-15 antigens are coexpressed on the same cells, and cell sorter-purified My4+ cells exhibit the morphology of monocytes. Whole blood analysis (which also shows staining of granulocytes) reveals that the average percentage of My4+ monocytes amongst all leucocytes is 5.8% with 580 cells/microliters. Furthermore, porcine peritoneal macrophages (PM) and alveolar macrophages (AM), both stain for My4, with a four-fold lower level on AM. Treatment of cells with phosphatidylinositol-specific phospholipase C decreases My4 staining, but does not affect staining with antibody 74-22-15. Immunoprecipitation with the My4 antibody from surface labelled pig mononuclear cells demonstrates a 54 kDa band similar to human CD14, and Western blotting with pig serum demonstrates two bands similar to the alpha and beta forms of human soluble CD14. Finally, the My4 antibody is capable of blocking lipopolysaccharide- (LPS)-induced interleukin-6 production in isolated PBMC. These data show that the My4 antibody recognizes genuine CD14 on porcine monocytes and macrophages.
已经制备了几种针对人CD14抗原的单克隆抗体。我们现在报告，抗体My4而非LeuM3可与猪单核细胞发生反应。在猪外周血单个核细胞（PBMC）中，14.6%的细胞可被CD14抗体My4染色，这与用抗猪单核细胞抗体74-22-15获得的百分比相似。双色免疫荧光显示My4和74-22-15抗原在同一细胞上共表达，且细胞分选纯化的My4+细胞呈现单核细胞的形态。全血分析（也显示粒细胞有染色）表明，所有白细胞中My4+单核细胞的平均百分比为5.8%，每微升有580个细胞。此外，猪腹腔巨噬细胞（PM）和肺泡巨噬细胞（AM）均对My4染色，AM上的染色水平低四倍。用磷脂酰肌醇特异性磷脂酶C处理细胞可降低My4染色，但不影响抗体74-22-15的染色。用My4抗体对表面标记的猪单核细胞进行免疫沉淀显示出一条与人类CD14相似的54 kDa条带，用猪血清进行蛋白质印迹显示出两条与人类可溶性CD14的α和β形式相似的条带。最后，My4抗体能够阻断分离的PBMC中脂多糖（LPS）诱导的白细胞介素-6的产生。这些数据表明，My4抗体可识别猪单核细胞和巨噬细胞上的真正CD14。
