title: Evaluation of influenza-specific humoral response by microbead array analysis.
abstract: BACKGROUND
  Quantitation and determination of antigen specificity of systemic and mucosal immune responses to influenza vaccination is beneficial for future vaccine development. Previous methods to acquire this information were costly, time consuming and sample exhaustive. The benefits of suspension microbead array (MBA) analysis are numerous. The multiplex capabilities of the system conserve time, money and sample, while generating statistically powerful data.
  
  
  OBJECTIVE
  To demonstrate the use of the assay by comparing the humoral influenza-specific responses of two cohorts from two countries that differed in circulating influenza strains and rates of influenza vaccination.
  
  
  METHODS
  Influenza hemagglutinin (HA) from different strains were coated on microbeads and incubated with serum samples to capture immunoglobulin (Ig) A(1) and IgG(1) host antibodies.
  
  
  RESULTS
  Statistically significant differences in IgA(1) and IgG(1) exist between the serum samples from Winnipeg (Manitoba) donors and those from Kenyan (Africa) donors. Data were compared using Mann-Whitney nonparametric tests. The Winnipeg donors had higher mean fluorescence intensity values, with significant P values for anti-HA IgA(1) to A/Wyoming/3/2003 (P=0.044), A/Vietnam/1203/2004 (P=0.0179), A/New Caledonia/20/99 (P<0.0001) and B/Tokyo/53/99 (P=0.0002). No differences were seen between the groups in their response to B/Jilin/20/2003. The Winnipeg donors had higher mean fluorescence intensity values, with significant P values for anti-HA IgG(1) to A/Wyoming/3/2003 (P=0.0135), B/Tokyo/53/99 (P=0.006) and B/Jilin20/2003 (P=0.026).
  
  
  CONCLUSIONS
  Influenza-specific IgA(1) and IgG(1) antibodies were successfully detected using MBA technology. A significant difference in antibody response was observed between Winnipeg and Kenyan donor serums. MBA analysis is a relatively quick and cost-effective method for serum antibody analysis. The potential to simultaneously assay small sample volumes for a multitude of antigens makes this method invaluable for future vaccine response monitoring.
authors.affiliation: Department of Medical Microbiology, University of Manitoba, Winnipeg, Manitoba.
authors.forename: Yoav
authors.initials: Y
authors.lastname: Keynan
authors.forename: Tavis
authors.initials: T
authors.lastname: Bodnarchuk
authors.forename: Stephen
authors.initials: S
authors.lastname: Wayne
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Keith R
authors.initials: KR
authors.lastname: Fowke