title: Effect of the β-propiolactone treatment on the adsorption and fusion of influenza A/Brisbane/59/2007 and A/New Caledonia/20/1999 virus H1N1 on a dimyristoylphosphatidylcholine/ganglioside GM3 mixed phospholipids monolayer at the air-water interface.
abstract: The production protocol of many whole cell/virion vaccines involves an inactivation step with β-propiolactone (BPL). Despite the widespread use of BPL, its mechanism of action is poorly understood. Earlier work demonstrated that BPL alkylates nucleotide bases, but its interaction with proteins has not been studied in depth. In the present study we use ellipsometry to analyze the influence of BPL treatment of two H1N1 influenza strains, A/Brisbane/59/2007 and A/New Caledonia/20/1999, which are used for vaccine production on an industrial scale. Analyses were conducted using a mixed lipid monolayer containing ganglioside GM3, which functions as the viral receptor. Our results show that BPL treatment of both strains reduces viral affinity for the mixed monolayer and also diminishes the capacity of viral domains to self-assemble. In another series of experiments, the pH of the subphase was reduced from 7.4 to 5 to provoke the pH-induced conformational change of hemagglutinin, which occurs following endocytosis into the endosome. In the presence of the native virus the pH decrease caused a reduction in domain size, whereas lipid layer thickness and surface pressure were increased. These observations are consistent with a fusion of the viral membrane with the lipid monolayer. Importantly, this fusion was not observed with adsorbed inactivated virus, which indicates that BPL treatment inhibits the first step of virus-membrane fusion. Our data also indicate that BPL chemically modifies hemagglutinin, which mediates the interaction with GM3.
authors: CBMN, UMR CNRS 5248, Université Bordeaux, IPB, Allée Geoffroy Saint Hilaire 33600 Pessac, France. b.desbat@cbmn.u-bordeaux.fr
authors: Bernard
authors: B
authors: Desbat
authors: Eloïse
authors: E
authors: Lancelot
authors: Tino
authors: T
authors: Krell
authors: Marie-Claire
authors: MC
authors: Nicolaï
authors: Fred
authors: F
authors: Vogel
authors: Michel
authors: M
authors: Chevalier
authors: Frédéric
authors: F
authors: Ronzon
mesh_terms: D000327:Adsorption; D000388:Air; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry*; D004705:Endocytosis; D005679:G(M3) Ganglioside / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D053118:Influenza A Virus, H1N1 Subtype / Q000187:drug effects* / Q000502:physiology; D011420:Propiolactone / Q000494:pharmacology*; D014867:Water
chemical_list: D005679:G(M3) Ganglioside; D014867:Water; D011420:Propiolactone; D004134:Dimyristoylphosphatidylcholine