Human interferon alpha enters cells by receptor-mediated endocytosis.
A small number of Escherichia coli-derived human interferon alpha A molecules (up to approximately 800/cell) bind specifically to high-affinity cell surface receptors on bovine kidney cells at 4 degrees. When the cells are subsequently warmed to 37 degrees, the amount of surface-bound interferon alpha (IFN-alpha) as recognized by a radiolabeled monoclonal antibody rapidly decreases. Using 125I-IFN-alpha and treatment of cells with acetic acid/sodium chloride to remove surface-bound IFN, a similar decrease of surface-bound IFN and a corresponding increase in intracellular radiolabeled material is observed following the temperature shift. An analysis of the trichloroacetic acid precipitability of the radiolabeled material in the medium, removed by acid treatment or internalized, shows that IFN is degraded intracellularly and that its degradation products are then released into the medium. The process of uptake, degradation, and release of degraded material can be inhibited by the lysosomotropic agent chloroquine. A stable conjugate of IFN with 5-nm colloidal gold was prepared without a detectable loss of antiviral activity. As shown by transmission electron microscopy, the conjugate, bound to cells at 4 degrees, was found in clathrin-coated pits and later in receptosomes following a temperature shift to 37 degrees. Morphometric quantitation showed that an excess of native IFN added during binding of the conjugate in the cold reduced the appearance of conjugate in receptosomes by 80%. These studies demonstrate that at least a portion of receptor-bound IFN enters cells by receptor-mediated endocytosis.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000502:physiology; D001666:Binding Sites, Antibody; D002417:Cattle; D002478:Cells, Cultured; D004705:Endocytosis; D004926:Escherichia coli; D006801:Humans; D007370:Interferon Type I / Q000378:metabolism*; D007668:Kidney; D008024:Ligands; D008854:Microscopy, Electron; D011956:Receptors, Cell Surface / Q000378:metabolism*; D017471:Receptors, Interferon; D013696:Temperature
D000911:Antibodies, Monoclonal; D007370:Interferon Type I; D008024:Ligands; D011956:Receptors, Cell Surface; D017471:Receptors, Interferon