title: Comparison of Pandemrix and Arepanrix, two pH1N1 AS03-adjuvanted vaccines differentially associated with narcolepsy development.
abstract: Narcolepsy onset in children has been associated with the 2009 influenza A H1N1 pandemic and vaccination with Pandemrix. However it was not clearly observed with other adjuvanted pH1N1 vaccines such as Arepanrix or Focetria. Our aim was to characterize the differences between Pandemrix and Arepanrix that might explain the risk for narcolepsy after Pandemrix vaccination using 2D-DIGE and mass spectrometry (MS). We found that Pandemrix (2009 batch) and Arepanrix (2010 batch) showed 5 main viral proteins: hemagglutinin HA1 and HA2 subunits, neuraminidase NA, nucleoprotein NP, and matrix protein MA1 and non-viral proteins from the Gallus gallus growth matrix used in the manufacturing of the vaccines. Latticed patterns of HA1, HA2 and NA indicated charge and molecular weight heterogeneity, a phenomenon likely caused by glycosylation and sulfation. Overall, Pandemrix contained more NP and NA, while Arepanrix displayed a larger diversity of viral and chicken proteins, with the exception of five chicken proteins (PDCD6IP, TSPAN8, H-FABP, HSP and TUB proteins) that were relatively more abundant in Pandemrix. Glycosylation patterns were similar in both vaccines. A higher degree of deamidation and dioxidation was found in Pandemrix, probably reflecting differential degradation across batches. Interestingly, HA1 146N (residue 129N in the mature protein) displayed a 10-fold higher deamidation in Arepanrix versus Pandemrix. In recent vaccine strains and Focetria, 146N is mutated to D which is associated with increased production yields suggesting that 146N deamidation may have also occurred during the manufacturing of Arepanrix. The presence of 146N in large relative amounts in Pandemrix and the wild type virus and in lower relative quantities in Arepanrix or other H1N1 vaccines may have affected predisposition to narcolepsy.
authors.affiliation: Center for Sleep Sciences and Medicine, Stanford School of Medicine, Palo Alto, CA, USA.
authors.forename: Louis
authors.initials: L
authors.lastname: Jacob
authors.affiliation: Stanford University Mass Spectrometry, Palo Alto, CA, USA.
authors.forename: Ryan
authors.initials: R
authors.lastname: Leib
authors.affiliation: Center for Sleep Sciences and Medicine, Stanford School of Medicine, Palo Alto, CA, USA.
authors.forename: Hanna M
authors.initials: HM
authors.lastname: Ollila
authors.affiliation: Center for Sleep Sciences and Medicine, Stanford School of Medicine, Palo Alto, CA, USA.
authors.forename: Mélodie
authors.initials: M
authors.lastname: Bonvalet
authors.affiliation: Stanford University Mass Spectrometry, Palo Alto, CA, USA.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Adams
authors.affiliation: Center for Sleep Sciences and Medicine, Stanford School of Medicine, Palo Alto, CA, USA. Electronic address: mignot@stanford.edu.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Mignot
mesh_terms: D006801:Humans; D007252:Influenza Vaccines / Q000009:adverse effects / Q000378:metabolism*; D007251:Influenza, Human / Q000517:prevention & control; D013058:Mass Spectrometry; D009290:Narcolepsy / Q000209:etiology*; D014611:Vaccination / Q000009:adverse effects*; D014764:Viral Proteins / Q000378:metabolism*
chemical_list: D007252:Influenza Vaccines; D014764:Viral Proteins; C568072:arepanrix; C556153:pandemrix
keywords: 2D-DIGE; Arepanrix; H1N1; Influenza A; Mass spectrometry; Narcolepsy; Pandemrix; Vaccine