Cytoplasmic as opposed to surface Ia antigens expressed on human peripheral blood lymphocytes and monocytes

Peripheral blood leucocytes were examined for the presence of HLA-Dr or Ia-like antigen on the cell surface and in the cytoplasm. Surface staining of viable cell suspensions, using monoclonal anti-Ia antibody in both immunofluorescence and immunoperoxidase methods gave similar results. Using a newly developed immunoperoxidase double staining procedure, which stains the cells when either viable or fixed, both the surface and cytoplasm of the individual cells were labelled and examined. Twenty to thirty percent of the mononuclear cells were positive for Ia staining on the cell surface and in the cytoplasm. Morphologically these positive cells were identified as both lymphocytes and monocytes. A small percentage (5-9%) of these monocytes and lymphocytes consistently labelled only cytoplasmic Ia determinants. In contrast, only surface binding was observed with monoclonal antibodies recognizing T cell subpopulations. The ability to detect intracellular Ia antigenic determinants is of value in distinguishing stages of lymphocyte and monocyte development.