CD68 antigen expression by human retinal pigment epithelial cells

Abstract

Although a primary role of the retinal pigment epithelium (RPE) is the phagocytosis of aged outer segment membranes, the RPE may also phagocytize particulates via several specific receptors that are characteristically present on mononuclear phagocytes of bone marrow origin. In recent immunophenotypic studies, CD68 monoclonal antibodies (mAb) have been shown to react selectively with a specific 110 kDa cytoplasmic glycoprotein present in mononuclear phagocytes from various sources. Designated as anti-macrophage antibodies that react with this macrophage-associated antigen. CD68 antibodies are now widely used for immunohistochemical identification of mononuclear phagocytes. Using a panel of CD68 mAb (KP1, EBM11, Ki-M6, Y1/82A, and Y2/131) we performed immunohistochemistry on three cytospin preparations of freshly isolated human RPE cells, three primary human RPE cultures, and 12 human RPE cell lines maintained in culture for up to 40 passages. Cytospin preparations of freshly isolated RPE cells demonstrated heavy reactivity in 5% of cells. Five- to 7-day-old primary RPE cultures exhibited uniform, heavy staining of all cells. Strong immunohistochemical reactivity persisted in all 12 cell lines at various passages up to and including passage 40. Stimulation of cultured RPE cells with interferon-gamma (100 U ml-1) for 24 and 48 hr did not produce observable differences in CD68 staining. RPE cells failed to stain when control mAb or mouse serum were substituted for the primary antibody. The constitutive expression of CD68 by neuroectodermally-derived RPE cells extends their immunophenotypic similarities with mesenchymally-derived mononuclear phagocytes and provides an additional antigenic marker to identify RPE cells in vitro.