Antibodies to phospholipids and liposomes: binding of antibodies to cells.
Binding of two monoclonal anti-liposome antibodies to the surface of cultured murine peritoneal macrophages was investigated by indirect immunofluorescence and enzyme-linked immunosorbent assay. Neither antibody bound to cultures of freshly explanted, nonadherent macrophages, but immunoreactivity was observed following cell adherence to tissue culture plastic. Fluorescent microscopic evaluation revealed heterogeneity in staining patterns of the antibodies on adherent cells. Binding both to viable and fixed adherent macrophages was observed even after a 10,000-fold dilution of antibody. Treatment of adherent macrophage cultures with trypsin increased antibody binding. Further treatment of trypsinized-macrophages with alkaline phosphatase or neuraminidase did not affect antibody binding, but phospholipase D and, to a greater extent, phospholipase C resulted in a marked decrease in cellular binding. The data indicate that antibodies produced against liposomes appear to bind to surface phospholipids of macrophages, but binding can be influenced by the physiological state of the macrophage and overlying cell surface proteins.
Fogler
W E
WE
Department of Membrane Biochemistry, Walter Reed Army Institute of Research, Washington, DC 20307-5100.
Swartz
G M
GM
Alving
C R
CR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000954:Antigens, Surface / Q000276:immunology; D002448:Cell Adhesion; D002478:Cells, Cultured; D002784:Cholesterol / Q000276:immunology; D004134:Dimyristoylphosphatidylcholine / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D007075:Immunoglobulin M / Q000276:immunology; D008081:Liposomes / Q000276:immunology*; D008264:Macrophages / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D010755:Organophosphates / Q000276:immunology; D010529:Peritoneal Cavity / Q000166:cytology; D010716:Phosphatidylinositols / Q000276:immunology; D010739:Phospholipase D / Q000494:pharmacology; D010743:Phospholipids / Q000276:immunology*; D014357:Trypsin / Q000494:pharmacology; D010738:Type C Phospholipases / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D007075:Immunoglobulin M; D008081:Liposomes; D010755:Organophosphates; D010716:Phosphatidylinositols; D010743:Phospholipids; C002290:dicetylphosphate; D002784:Cholesterol; D010738:Type C Phospholipases; D010739:Phospholipase D; D014357:Trypsin; D004134:Dimyristoylphosphatidylcholine