title: Neutralization of influenza virus by low concentrations of hemagglutinin-specific polymeric immunoglobulin A inhibits viral fusion activity, but activation of the ribonucleoprotein is also inhibited.
abstract: High concentrations of hemagglutinin-specific neutralizing polymeric monoclonal immunoglobulin A (IgA) inhibit attachment of the majority of type A influenza virus virions to cell monolayers and tracheal epithelium (H. P. Taylor and N. J. Dimmock, J. Exp. Med. 161:198-209, 1985; M. C. Outlaw and N. J. Dimmock, J. Gen. Virol. 71:69-76, 1990). A minority of virions attaches but is not infectious. Here, we report that a different mechanism operates when influenza virus A/Puerto Rico/8/34 (H1N1) is neutralized by low concentrations of monoclonal polymeric IgA or when A/fowl plague virus/Rostock/34 (H7N1) is neutralized by low concentrations of polyclonal rat secretory IgA. Under these conditions, neutralized virus attaches to cells and is taken up by them. However, upon entering the cell, the nucleoprotein (NP) of neutralized virus is found in the perinuclear cytoplasm, whereas NP from nonneutralized virus is concentrated in the nucleus itself. Further data show that the low-pH-mediated cell fusion activity of virions is inhibited by IgA in proportion to loss of infectivity. The possibilities that neutralization by low amounts of polymeric IgA is caused by inhibition of the virion fusion activity and that the aberrant distribution of NP from neutralized virus results from its failure to escape from the endosomal system were investigated by using A/PR/8/34 and the fusogenic agent polyethylene glycol (PEG) at pH 5.4. A/PR/8/34 attached to cells at 4 degrees C, with minimal internalization of the virus; treatment with PEG at pH 5.4 and 4 degrees C for 1 min led to infectious fusion of nonneutralized virus with the plasma membrane and, under these conditions, was more efficient than PEG at pH 7 or medium at pH 5.4. Neutralized virus which was attached to cells and treated with acidified PEG appeared to undergo primary and secondary uncoating, with its NP protein becoming concentrated in the nucleus and M1 becoming concentrated in the perinuclear cytoplasm. Although the distribution of NP and M1 was indistinguishable from infectious virus, infectivity was not restored. Thus, even when IgA-induced inhibition of fusion is reversed, virus is still neutralized. We suggest that infectious influenza virus undergoes an activation stage which may be the relaxation of the ribonucleoprotein structure needed to permit transcription or may be the removal of M1 bound to the ribonucleoprotein.(ABSTRACT TRUNCATED AT 400 WORDS)
authors.affiliation: Department of Biological Sciences, University of Warwick, Coventry, United Kingdom.
authors.forename: S J
authors.initials: SJ
authors.lastname: Armstrong
authors.forename: N J
authors.initials: NJ
authors.lastname: Dimmock
mesh_terms: D000818:Animals; D001287:Attachment Sites, Microbiological / Q000187:drug effects / Q000276:immunology*; D002459:Cell Fusion / Q000187:drug effects; D002467:Cell Nucleus / Q000378:metabolism; D002642:Chick Embryo; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral / Q000276:immunology*; D007070:Immunoglobulin A / Q000276:immunology*; D009980:Influenza A virus / Q000276:immunology / Q000472:pathogenicity*; D009500:Neutralization Tests; D011092:Polyethylene Glycols / Q000494:pharmacology; D011991:Receptors, Virus / Q000378:metabolism; D012261:Ribonucleoproteins / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000276:immunology*; D014764:Viral Proteins / Q000378:metabolism; D014771:Virion / Q000378:metabolism; D014775:Virus Activation; D014779:Virus Replication
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral; D007070:Immunoglobulin A; D011991:Receptors, Virus; D012261:Ribonucleoproteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; D011092:Polyethylene Glycols