Koch A E, Burrows J C, Marder R, Domer P H, Leibovich S J
Department of Medicine, Northwestern University Medical School, Chicago, Ill.
Pathobiology. 1990;58(5):241-8. doi: 10.1159/000163594.
We have characterized antimyeloid monoclonal antibodies (mAbs) produced to human rheumatoid arthritis (RA) synovial tissue macrophages (MPs) (8D7) and to lipopolysaccharide (LPS)-treated U937 cells (3D8). The 3D8 antigen is upregulated with LPS stimulation of monocytes/MPs and during monocyte maturation. The 8D7 antigen is upregulated on functionally distinct subpopulations of RA synovial tissue MPs. We used immunohistochemistry to determine the spectrum of reactivity of these unique mAbs on myeloid cell suspensions, monocytes, and mature tissue inflammatory and noninflammatory MPs. The antigens identified by the mAbs were characterized biochemically, by immunoprecipitation of solubilized 125I-labelled antigens from cell surfaces, and immunohistochemically by enzymatic digestion of myeloid cells followed by a cellular ELISA. MAb 3D8, characterized as an anti-CD13 antibody, recognizes a 150-170 kd antigen, has almost exclusive myeloid reactivity, but reacts with Langerhans' cells of the skin and thymus, pointing to shared antigens between these cells and MPs. Unlike 3D8 antigen, 8D7 antigen is strongly expressed in inflammatory states, being present on MPs in granulomata as well as in sarcoid lymph nodes. Both mAbs react with frozen and methanol-Carnoy's fixed, paraffin-embedded tissues and detect antigenic differences among human mononuclear phagocytes present in different anatomical sites and in varying stages of differentiation and activation. These mAbs should prove to be a valuable tool for studying heterogenous populations of myeloid cells.
我们已经对针对人类类风湿性关节炎(RA)滑膜组织巨噬细胞(MPs)(8D7)以及脂多糖(LPS)处理的U937细胞(3D8)产生的抗髓系单克隆抗体(mAbs)进行了特性鉴定。3D8抗原在单核细胞/ MPs受到LPS刺激时以及单核细胞成熟过程中上调。8D7抗原在RA滑膜组织MPs功能不同的亚群上上调。我们使用免疫组织化学来确定这些独特的mAbs在髓系细胞悬液、单核细胞以及成熟组织炎性和非炎性MPs上的反应谱。通过从细胞表面溶解的125I标记抗原的免疫沉淀对mAbs识别的抗原进行生化特性鉴定,并通过对髓系细胞进行酶消化然后进行细胞ELISA进行免疫组织化学鉴定。特征为抗CD13抗体的mAb 3D8识别一种150 - 170 kd的抗原,几乎具有排他性的髓系反应性,但与皮肤和胸腺的朗格汉斯细胞反应,表明这些细胞与MPs之间存在共同抗原。与3D8抗原不同,8D7抗原在炎症状态下强烈表达,存在于肉芽肿以及结节病淋巴结的MPs上。两种mAbs都与冷冻以及甲醇 - 卡诺固定、石蜡包埋的组织反应,并检测存在于不同解剖部位以及不同分化和激活阶段的人类单核吞噬细胞之间的抗原差异。这些mAbs应该被证明是研究髓系细胞异质群体的有价值工具。