title: Characterization of influenza vaccine immunogenicity using influenza antigen microarrays.
abstract: BACKGROUND
  Existing methods to measure influenza vaccine immunogenicity prohibit detailed analysis of epitope determinants recognized by immunoglobulins. The development of highly multiplex proteomics platforms capable of capturing a high level of antibody binding information will enable researchers and clinicians to generate rapid and meaningful readouts of influenza-specific antibody reactivity.
  
  
  METHODS
  We developed influenza hemagglutinin (HA) whole-protein and peptide microarrays and validated that the arrays allow detection of specific antibody reactivity across a broad dynamic range using commercially available antibodies targeted to linear and conformational HA epitopes. We derived serum from blood draws taken from 76 young and elderly subjects immediately before and 28±7 days post-vaccination with the 2008/2009 trivalent influenza vaccine and determined the antibody reactivity of these sera to influenza array antigens.
  
  
  RESULTS
  Using linear regression and correcting for multiple hypothesis testing by the Benjamini and Hochberg method of permutations over 1000 resamplings, we identified antibody reactivity to influenza whole-protein and peptide array features that correlated significantly with age, H1N1, and B-strain post-vaccine titer as assessed through a standard microneutralization assay (p<0.05, q <0.2). Notably, we identified several peptide epitopes that were inversely correlated with regard to age and seasonal H1N1 and B-strain neutralization titer (p<0.05, q <0.2), implicating reactivity to these epitopes in age-related defects in response to H1N1 influenza. We also employed multivariate linear regression with cross-validation to build models based on age and pre-vaccine peptide reactivity that predicted vaccine-induced neutralization of seasonal H1N1 and H3N2 influenza strains with a high level of accuracy (84.7% and 74.0%, respectively).
  
  
  CONCLUSIONS
  Our methods provide powerful tools for rapid and accurate measurement of broad antibody-based immune responses to influenza, and may be useful in measuring response to other vaccines and infectious agents.
authors.affiliation: Department of Medicine, Division of Immunology and Rheumatology, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Jordan V
authors.initials: JV
authors.lastname: Price
authors.forename: Justin A
authors.initials: JA
authors.lastname: Jarrell
authors.forename: David
authors.initials: D
authors.lastname: Furman
authors.forename: Nicole H
authors.initials: NH
authors.lastname: Kattah
authors.forename: Evan
authors.initials: E
authors.lastname: Newell
authors.forename: Cornelia L
authors.initials: CL
authors.lastname: Dekker
authors.forename: Mark M
authors.initials: MM
authors.lastname: Davis
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Utz
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000595:Amino Acid Sequence; D000914:Antibodies, Viral / Q000276:immunology / Q000378:metabolism; D000956:Antigens, Viral / Q000276:immunology* / Q000378:metabolism; D000939:Epitopes / Q000276:immunology / Q000378:metabolism; D005260:Female; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology*; D053122:Influenza A Virus, H3N2 Subtype / Q000276:immunology*; D007252:Influenza Vaccines / Q000008:administration & dosage / Q000276:immunology*; D007251:Influenza, Human / Q000276:immunology* / Q000517:prevention & control / Q000821:virology; D016014:Linear Models; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology / Q000378:metabolism; D040081:Protein Array Analysis / Q000379:methods; D040901:Proteomics / Q000379:methods; D014611:Vaccination; D055815:Young Adult
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D000939:Epitopes; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007252:Influenza Vaccines; D010455:Peptides