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dc.contributor.authorde Silva, Thushan I
dc.contributor.authorGould, Victoria
dc.contributor.authorMohammed, Nuredin I
dc.contributor.authorCope, Alethea
dc.contributor.authorMeijer, Adam
dc.contributor.authorZutt, Ilse
dc.contributor.authorReimerink, Johan
dc.contributor.authorKampmann, Beate
dc.contributor.authorHoschler, Katja
dc.contributor.authorZambon, Maria
dc.contributor.authorTregoning, John S
dc.date.accessioned2018-01-02T14:11:52Z
dc.date.available2018-01-02T14:11:52Z
dc.date.issued2017-10
dc.identifier.citationComparison of mucosal lining fluid sampling methods and influenza-specific IgA detection assays for use in human studies of influenza immunity. 2017, 449:1-6 J. Immunol. Methodsen
dc.identifier.issn1872-7905
dc.identifier.pmid28647455
dc.identifier.doi10.1016/j.jim.2017.06.008
dc.identifier.urihttp://hdl.handle.net/10029/620966
dc.description.abstractWe need greater understanding of the mechanisms underlying protection against influenza virus to develop more effective vaccines. To do this, we need better, more reproducible methods of sampling the nasal mucosa. The aim of the current study was to compare levels of influenza virus A subtype-specific IgA collected using three different methods of nasal sampling. Samples were collected from healthy adult volunteers before and after LAIV immunization by nasal wash, flocked swabs and Synthetic Absorptive Matrix (SAM) strips. Influenza A virus subtype-specific IgA levels were measured by haemagglutinin binding ELISA or haemagglutinin binding microarray and the functional response was assessed by microneutralization. Nasosorption using SAM strips lead to the recovery of a more concentrated sample of material, with a significantly higher level of total and influenza H1-specific IgA. However, an equivalent percentage of specific IgA was observed with all sampling methods when normalized to the total IgA. Responses measured using a recently developed antibody microarray platform, which allows evaluation of binding to multiple influenza strains simultaneously with small sample volumes, were compared to ELISA. There was a good correlation between ELISA and microarray values. Material recovered from SAM strips was weakly neutralizing when used in an in vitro assay, with a modest correlation between the level of IgA measured by ELISA and neutralization, but a greater correlation between microarray-measured IgA and neutralizing activity. In conclusion we have tested three different methods of nasal sampling and show that flocked swabs and novel SAM strips are appropriate alternatives to traditional nasal washes for assessment of mucosal influenza humoral immunity.
dc.language.isoenen
dc.rightsArchived with thanks to Journal of immunological methodsen
dc.subject.meshAdult
dc.subject.meshAntibodies, Viral
dc.subject.meshEnzyme-Linked Immunosorbent Assay
dc.subject.meshFemale
dc.subject.meshHumans
dc.subject.meshImmunity, Mucosal
dc.subject.meshImmunoglobulin A
dc.subject.meshInfluenza A virus
dc.subject.meshInfluenza Vaccines
dc.subject.meshMale
dc.subject.meshNasal Lavage Fluid
dc.subject.meshNasal Mucosa
dc.subject.meshProtein Array Analysis
dc.subject.meshSpecimen Handling
dc.titleComparison of mucosal lining fluid sampling methods and influenza-specific IgA detection assays for use in human studies of influenza immunity.en
dc.typeArticleen
dc.identifier.journalJ Immunol Methods 2017, 449:1-6en
html.description.abstractWe need greater understanding of the mechanisms underlying protection against influenza virus to develop more effective vaccines. To do this, we need better, more reproducible methods of sampling the nasal mucosa. The aim of the current study was to compare levels of influenza virus A subtype-specific IgA collected using three different methods of nasal sampling. Samples were collected from healthy adult volunteers before and after LAIV immunization by nasal wash, flocked swabs and Synthetic Absorptive Matrix (SAM) strips. Influenza A virus subtype-specific IgA levels were measured by haemagglutinin binding ELISA or haemagglutinin binding microarray and the functional response was assessed by microneutralization. Nasosorption using SAM strips lead to the recovery of a more concentrated sample of material, with a significantly higher level of total and influenza H1-specific IgA. However, an equivalent percentage of specific IgA was observed with all sampling methods when normalized to the total IgA. Responses measured using a recently developed antibody microarray platform, which allows evaluation of binding to multiple influenza strains simultaneously with small sample volumes, were compared to ELISA. There was a good correlation between ELISA and microarray values. Material recovered from SAM strips was weakly neutralizing when used in an in vitro assay, with a modest correlation between the level of IgA measured by ELISA and neutralization, but a greater correlation between microarray-measured IgA and neutralizing activity. In conclusion we have tested three different methods of nasal sampling and show that flocked swabs and novel SAM strips are appropriate alternatives to traditional nasal washes for assessment of mucosal influenza humoral immunity.


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