title: Antigenic differences between AS03 adjuvanted influenza A (H1N1) pandemic vaccines: implications for pandemrix-associated narcolepsy risk.
abstract: BACKGROUND
  Narcolepsy results from immune-mediated destruction of hypocretin secreting neurons in hypothalamus, however the triggers and disease mechanisms are poorly understood. Vaccine-attributable risk of narcolepsy reported so far with the AS03 adjuvanted H1N1 vaccination Pandemrix has been manifold compared to the AS03 adjuvanted Arepanrix, which contained differently produced H1N1 viral antigen preparation. Hence, antigenic differences and antibody response to these vaccines were investigated.
  
  
  RESULTS
  Increased circulating IgG-antibody levels to Pandemrix H1N1 antigen were found in 47 children with Pandemrix-associated narcolepsy when compared to 57 healthy children vaccinated with Pandemrix. H1N1 antigen of Arepanrix inhibited poorly these antibodies indicating antigenic difference between Arepanrix and Pandemrix. High-resolution gel electrophoresis quantitation and mass spectrometry identification analyses revealed higher amounts of structurally altered viral nucleoprotein (NP) in Pandemrix. Increased antibody levels to hemagglutinin (HA) and NP, particularly to detergent treated NP, was seen in narcolepsy. Higher levels of antibodies to NP were found in children with DQB1*06:02 risk allele and in DQB1*06:02 transgenic mice immunized with Pandemrix when compared to controls.
  
  
  CONCLUSIONS
  This work identified 1) higher amounts of structurally altered viral NP in Pandemrix than in Arepanrix, 2) detergent-induced antigenic changes of viral NP, that are recognized by antibodies from children with narcolepsy, and 3) increased antibody response to NP in association of DQB1*06:02 risk allele of narcolepsy. These findings provide a link between Pandemrix and narcolepsy. Although detailed mechanisms of Pandemrix in narcolepsy remain elusive, our results move the focus from adjuvant(s) onto the H1N1 viral proteins.
authors.affiliation: Department of Vaccinations and Immune Protection, National Institute for Health and Welfare, Helsinki, Finland.
authors.forename: Outi
authors.initials: O
authors.lastname: Vaarala
authors.affiliation: Department of Vaccinations and Immune Protection, National Institute for Health and Welfare, Helsinki, Finland.
authors.forename: Arja
authors.initials: A
authors.lastname: Vuorela
authors.affiliation: Helsinki Sleep Clinic, Vitalmed Research Centre Helsinki and Haartman Institute, University of Helsinki, Helsinki, Finland.
authors.forename: Markku
authors.initials: M
authors.lastname: Partinen
authors.affiliation: Meilahti Clinical Proteomics Core Facility, Institute of Biomedicine/Biochemistry and Developmental Biology, and NeuroMed Research Program, University of Helsinki, Helsinki, Finland.
authors.forename: Marc
authors.initials: M
authors.lastname: Baumann
authors.affiliation: Department of Bacteriology and Immunology, Haartman Institute, University of Helsinki, Helsinki, Finland and Research Program Unit, Immunobiology, University of Helsinki, Helsinki, Finland.
authors.forename: Tobias L
authors.initials: TL
authors.lastname: Freitag
authors.affiliation: Department of Bacteriology and Immunology, Haartman Institute, University of Helsinki, Helsinki, Finland and Research Program Unit, Immunobiology, University of Helsinki, Helsinki, Finland.
authors.forename: Seppo
authors.initials: S
authors.lastname: Meri
authors.affiliation: Department of Bacteriology and Immunology, Haartman Institute, University of Helsinki, Helsinki, Finland and Research Program Unit, Immunobiology, University of Helsinki, Helsinki, Finland.
authors.forename: Päivi
authors.initials: P
authors.lastname: Saavalainen
authors.affiliation: National Institute for Health and Welfare, Public Health Genomics Research Unit, Biomedicum, Helsinki, Finland.
authors.forename: Matti
authors.initials: M
authors.lastname: Jauhiainen
authors.affiliation: Meilahti Clinical Proteomics Core Facility, Institute of Biomedicine/Biochemistry and Developmental Biology, and NeuroMed Research Program, University of Helsinki, Helsinki, Finland.
authors.forename: Rabah
authors.initials: R
authors.lastname: Soliymani
authors.affiliation: Department of Pediatrics, Children's Hospital, Helsinki University Hospital, Helsinki, Finland.
authors.forename: Turkka
authors.initials: T
authors.lastname: Kirjavainen
authors.affiliation: Department of Child Neurology, Oulu University Hospital, Oulu, Finland.
authors.forename: Päivi
authors.initials: P
authors.lastname: Olsen
authors.affiliation: Department of Pediatrics, Tampere University Hospital, Tampere, Finland.
authors.forename: Outi
authors.initials: O
authors.lastname: Saarenpää-Heikkilä
authors.affiliation: Department of Chemistry and Biocenter Kuopio, University of Eastern Finland, Joensuu, Finland.
authors.forename: Juha
authors.initials: J
authors.lastname: Rouvinen
authors.affiliation: Department of Infectious Disease Surveillance and Control, National Institute for Health and Welfare, Helsinki, Finland.
authors.forename: Merja
authors.initials: M
authors.lastname: Roivainen
authors.affiliation: Department of Vaccinations and Immune Protection, National Institute for Health and Welfare, Helsinki, Finland.
authors.forename: Hanna
authors.initials: H
authors.lastname: Nohynek
authors.affiliation: Department of Vaccinations and Immune Protection, National Institute for Health and Welfare, Helsinki, Finland.
authors.forename: Jukka
authors.initials: J
authors.lastname: Jokinen
authors.affiliation: Department of Infectious Disease Surveillance and Control, National Institute for Health and Welfare, Helsinki, Finland; Department of Virology, University of Turku, Turku, Finland.
authors.forename: Ilkka
authors.initials: I
authors.lastname: Julkunen
authors.affiliation: Department of Vaccinations and Immune Protection, National Institute for Health and Welfare, Helsinki, Finland.
authors.forename: Terhi
authors.initials: T
authors.lastname: Kilpi
mesh_terms: D000293:Adolescent; D000483:Alleles; D000818:Animals; D000917:Antibody Formation; D000956:Antigens, Viral / Q000009:adverse effects / Q000032:analysis / Q000276:immunology; D002648:Child; D002675:Child, Preschool; D059866:HLA-DQ beta-Chains / Q000235:genetics / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology*; D007252:Influenza Vaccines / Q000009:adverse effects* / Q000032:analysis / Q000276:immunology; D007251:Influenza, Human / Q000235:genetics / Q000276:immunology / Q000517:prevention & control*; D016688:Mice, Inbred NOD; D009290:Narcolepsy / Q000209:etiology* / Q000235:genetics / Q000276:immunology; D009976:Orthomyxoviridae Infections / Q000235:genetics / Q000276:immunology / Q000517:prevention & control
chemical_list: D000956:Antigens, Viral; D059866:HLA-DQ beta-Chains; C069051:HLA-DQB1 antigen; D007074:Immunoglobulin G; D007252:Influenza Vaccines; C568072:arepanrix; C556153:pandemrix