Hemolytic activity of influenza virus hemagglutinin glycoproteins activated in mildly acidic environments.
Hemagglutinin (HA) glycoproteins isolated from influenza virus caused hemolysis and liposome lysis at pH less than 6.0. The pH dependence was similar to that of the parent virus. Hemagglutination and hemolysis titers of HA were comparable with those of virus. The time course of hemolysis by HA was somewhat different from that by virus. HA did not cause fusion of erythrocytes in acidic media, in contrast to virus. Both HA and virus, previously incubated at pH less than 6.0, lost their low-pH-induced hemolytic activity. Isolated HA formed rosette-like structures at neutral pH, and these aggregated in acidic media. Virus also aggregated in acidic media and its envelope became leaky to negative stain. HA previously incubated at pH less than 6.0 became susceptible to trypsin digestion. Both reversible and irreversible structural changes of HA were observed by fluorescence spectroscopy; a reversible change at a pH between neutral and 6.4 and an irreversible one at pH less than 6.0. Bromelain-released HA did not cause hemolysis and liposome lysis in acidic media. The precursor form of HA did not have hemolytic activity in acidic media. The similarity in pH dependence indicates that the structural change in HA induced at pH less than 6.0 is the cause of activation and inactivation of hemolysis, HA and virus aggregation, and trypsin susceptibility. We propose that the hydrophobic NH2-terminal segment of HA2 is exposed during the structural change and interacts with the target membranes, causing a permeability increase and leading to hemolysis and lysis. The virus-induced hemolysis can be ascribed for the most part to envelope fusion activated in acidic media.
D000818:Animals; D002642:Chick Embryo; D006388:Hemagglutinins / Q000276:immunology*; D006461:Hemolysis; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009980:Influenza A virus / Q000276:immunology* / Q000648:ultrastructure; D007700:Kinetics; D008081:Liposomes; D008854:Microscopy, Electron
D006388:Hemagglutinins; D008081:Liposomes