Phagocytosis of antigens by Langerhans cells in vitro

Abstract

Dendritic cells (DC) isolated from lymphoid tissues are generally thought to be nonphagocytic in culture. It has therefore been unclear how these cells could acquire particulate antigens such as microorganisms for initiation of primary immune responses. Lymphoid DC derive in part from cells that have migrated from nonlymphoid tissues, such as Langerhans cells (LC) of skin. The ability of LC to internalize a variety of particles was studied by electron, ultraviolet, phase, and differential interference contrast microscopy, and by two-color flow cytometry. Freshly isolated LC in epidermal cell suspensions phagocytosed the yeast cell wall derivative zymosan, intact Saccharomyces cerevisiae, representatives of two genera of Gram-positive bacteria, Corynebacterium parvum and Staphylococcus aureus, as well as 0.5-3.5-microns latex microspheres. During maturation in culture, the phagocytic activity of these cells was markedly reduced. Likewise, freshly isolated splenic DC were more phagocytic than cultured DC for two types of particle examined, zymosan and latex beads. Unlike macrophages, LC did not bind or internalize sheep erythrocytes before or after opsonization with immunoglobulin G or complement, and did not internalize colloidal carbon. The receptors mediating zymosan uptake by LC were examined. For this particle, C57BL/6 LC were considerably more phagocytic than BALB/c LC and exhibited a reproducible increase in phagocytic activity after 6 h of culture followed by a decline, whereas this initial rise did not occur for BALB/c LC. These differential kinetics of uptake were reflected in the pattern of zymosan binding at 4 degrees C, and endocytosis of the soluble tracer fluorescein isothiocyanate-mannose-bovine serum albumin at 37 degrees C. Zymosan uptake by LC from both strains of mice was inhibited in the presence of mannan or beta-glucan, although to different extents, but not by antibodies specific for CR3 (CD11b/CD18). These data indicate that zymosan uptake by LC can be mediated by a mannose/beta-glucan receptor(s) that is differentially expressed in the two strains of mice and that is downregulated during maturation of LC in culture.