Characterization of influenza A virus binding sites on human neutrophils.
Exposure of human neutrophils (PMN) to influenza A virus (IAV) triggers discrete responses in these cells that interfere with their normal host defense functions. Because the restricted host range and tissue specificities of many viruses are determined by cell surface molecules acting as virus receptors on target cells, it seemed plausible that IAV might interact with neutrophils via specific plasma membrane glycoproteins that bind to viral hemagglutinin. When the binding of intact IAV (ATCC strain A/PR/8/34 (H1N1)) to PMNs was examined by flow cytometry, virus binding was found to be saturable and to be diminished after extensive desialation of the cells with neuraminidase. Stimulation of PMNs with FMLP (0.1 microM) caused a transient increase in IAV binding that was maximal (> 200%) at 2 min after stimulation. When neutrophil membrane proteins were separated by gel electrophoresis and transferred to nitrocellulose, IAV bound selectively to two polypeptide bands of approximately 125 and 160 kDa. Relative binding to these two bands was modified and ultimately eliminated by treatment of PMN membrane proteins with neuraminidase before electrophoresis and blotting. Intact virus precipitated a limited number of proteins from solubilized PMN plasma membrane preparations, and Abs specific for sialophorin (CD43) recognized virus-precipitated PMN membrane proteins of the same apparent m.w. as those detected in virus-membrane protein blots. These findings indicate that IAV binds to human PMNs through interactions with a limited number of PMN membrane glycoproteins, which include sialophorin (CD43).
Rothwell
S W
SW
Department of Hematology, Walter Reed Army Institute of Research, Washington, DC 20307-5100.
Wright
D G
DG
D015703:Antigens, CD / Q000378:metabolism; D001665:Binding Sites; D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism / Q000382:microbiology; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D066298:In Vitro Techniques; D009980:Influenza A virus / Q000276:immunology / Q000378:metabolism*; D051917:Leukosialin; D008562:Membrane Glycoproteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism / Q000382:microbiology*; D011991:Receptors, Virus / Q000276:immunology / Q000378:metabolism*; D012795:Sialoglycoproteins / Q000276:immunology / Q000378:metabolism
D015703:Antigens, CD; D051917:Leukosialin; D008562:Membrane Glycoproteins; D011991:Receptors, Virus; C465014:SPN protein, human; D012795:Sialoglycoproteins; D006861:Hydrogen Peroxide