Austyn J M, Gordon S
Eur J Immunol. 1981 Oct;11(10):805-15. doi: 10.1002/eji.1830111013.
A hybridoma clone which secretes a macrophage (M phi)-specific monoclonal antibody, F4/80, was produced by fusing spleen cells from a rat hyperimmunized with cultured thioglycollate-induced mouse peritoneal M phi with a mouse myeloma, NS1. Binding of antibody to primary cells and cell lines was detected by radioimmune indirect binding assay, autoradiography or fluorescence-activated cell sorter analysis. F4/80 binds to mouse M phi from the peritoneal cavity or other sources, blood monocytes, M phi derived from bone marrow precursors in culture and M phi-like cell lines, but not to other cells, including polymorphonuclear leukocytes, lymphocytes or fibroblasts. F4/80 does not bind to M phi via Fc receptors, is not cytotoxic and is of the rat IgG2b subclass. Since F4/80 binds to all M phi defined by adherence, morphology and immune phagocytosis, it provides a new marker to define the M phi in the mouse. Large differences in expression of antigen F4/80 were found, depending on intraperitoneal stimulation, time in culture and stage of maturation. Immunoprecipitation experiments demonstrated that the antigen F4/80 is part of a component of Mr 160000 which is synthesized by the M phi and, at least in part, exposed on the cell surface.
通过将用培养的巯基乙酸盐诱导的小鼠腹腔巨噬细胞超免疫的大鼠的脾细胞与小鼠骨髓瘤NS1融合,产生了一种分泌巨噬细胞(M phi)特异性单克隆抗体F4/80的杂交瘤克隆。通过放射免疫间接结合试验、放射自显影或荧光激活细胞分选分析检测抗体与原代细胞和细胞系的结合。F4/80与来自腹腔或其他来源的小鼠巨噬细胞、血液单核细胞、培养中源自骨髓前体的巨噬细胞以及巨噬细胞样细胞系结合,但不与其他细胞结合,包括多形核白细胞、淋巴细胞或成纤维细胞。F4/80不通过Fc受体与巨噬细胞结合,无细胞毒性,属于大鼠IgG2b亚类。由于F4/80与通过贴壁、形态和免疫吞噬作用定义的所有巨噬细胞结合,它为定义小鼠中的巨噬细胞提供了一种新的标志物。根据腹腔刺激、培养时间和成熟阶段的不同,发现抗原F4/80的表达存在很大差异。免疫沉淀实验表明,抗原F4/80是分子量为160000的一种成分的一部分,该成分由巨噬细胞合成,至少部分暴露在细胞表面。