Reactivity of Human Tissues with Monoclonal Antibodies to Myeloid Activation and Differentiation Antigens

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Abstract
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-CD 13 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 3D 8 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.

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