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A novel whole blood assay to quantify the release of T cell associated cytokines in response to Bordetella pertussis antigens

2024-11Contributo em revista Acesso aberto
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datacite.subject.fosCiências Médicas::Ciências da Saúde
datacite.subject.sdg03:Saúde de Qualidade
dc.contributor.authorPinto, Marta Valente
dc.contributor.authorBarkoff, Alex-Mikael
dc.contributor.authorBibi, Sagida
dc.contributor.authorKnuutila, Aapo
dc.contributor.authorTeräsjärvi, Johanna
dc.contributor.authorClutterbuck, Elizabeth
dc.contributor.authorGimenez-Fourage, Sophie
dc.contributor.authorPagnon, Anke
dc.contributor.authorGaans-van den Brink, Jacqueline A.M. van
dc.contributor.authorCorbiere, Veronique
dc.contributor.authorMontfort, Aymeric De
dc.contributor.authorSaso, Anja
dc.contributor.authorJobe, Haddijatou
dc.contributor.authorRoetynck, Sophie
dc.contributor.authorKampmann, Beate
dc.contributor.authorSimonetti, Elles
dc.contributor.authorDiavatopoulos, Dimitri
dc.contributor.authorLambert, Eleonora E.
dc.contributor.authorMertsola, Jussi
dc.contributor.authorBlanc, Pascal
dc.contributor.authorEls, Cécile A.C.M. van
dc.contributor.authorKelly, Dominic
dc.contributor.authorHe, Qiushui
dc.contributor.authorThe PERISCOPE Consortium
dc.date.accessioned2026-03-18T12:47:48Z
dc.date.available2026-03-18T12:47:48Z
dc.date.issued2024-11
dc.description.abstractBackground: Bordetella pertussis continues to cause whooping cough globally even in countries with high immunisation coverage. Booster vaccinations with acellular pertussis vaccines are thus used in children, adolescents, and adults. T cell immunity is crucial for orchestrating the immune response after vaccination. However, T cell assays can be expensive and difficult to implement in large clinical trials. In this study, a whole blood (WB) stimulation assay was developed to identify secreted T cell associated cytokines in different age groups after acellular pertussis booster vaccination. Material and methods: Longitudinal WB samples were collected from a small set of subjects (n = 38) aged 7–70 years participating in a larger ongoing clinical trial. For assay development, samples were diluted and incubated with purified inactivated pertussis toxin (PT), filamentous haemagglutinin (FHA), inactivated B. pertussis lysate, and complete medium (M) as stimulating conditions, with anti-CD28 and anti-CD49d as co-stimulants. Different timepoints around the vaccination (D0, D7, D14, D28), WB dilution factor (1:2, 1:4) and incubation time (24 h, 48 h, 72 h) were compared. Responses to 15 cytokines were tested with Luminex/multiplex immunoassay. Results: The optimized assay consisted of WB incubation with M, PT, and FHA (including the two co-stimulants). After 48 h incubation, supernatants were collected for measurement of seven selected T cell associated cytokines (IL-2, IL-5, IL-10, IL-13, IL-17 A, IL-17F, and IFN-y) from samples before and 28 days after vaccination. PT stimulation showed a trend for upregulation of IL-2, IL-13, and IL-17 A/F for adult subjects, whereas the responses of all cytokines were downregulated for the paediatric subjects. Furthermore, PT and FHA-stimulated WB showed diverse cytokine producing profiles. Conclusions: The developed WB-based cytokine assay was shown to be less costly, easy to perform, and functional in differently aged individuals. Further, it requires only a small amount of fresh blood, which is beneficial especially for studies including infants. Our results support the use of this assay for other immunological studies in the future.eng
dc.identifier.citationPinto, M. V., Barkoff, A. M., Bibi, S., Knuutila, A., Teräsjärvi, J., Clutterbuck, E., Gimenez-Fourage, S., Pagnon, A., van Gaans-van den Brink, J. A. M., Corbiere, V., De Montfort, A., Saso, A., Jobe, H., Roetynck, S., Kampmann, B., Simonetti, E., Diavatopoulos, D., Lambert, E. E., Mertsola, J., Blanc, P., … PERISCOPE Consortium (2024). A novel whole blood assay to quantify the release of T cell associated cytokines in response to Bordetella pertussis antigens. Journal of immunological methods, 534, 113758. https://doi.org/10.1016/j.jim.2024.113758
dc.identifier.doi10.1016/j.jim.2024.113758
dc.identifier.issn0022-1759
dc.identifier.urihttp://hdl.handle.net/10400.26/62267
dc.language.isoeng
dc.peerreviewedyes
dc.publisherElsevier
dc.relation.hasversionhttps://doi.org/10.1016/j.jim.2024.113758
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectBordetella pertussis
dc.subjectCytokines
dc.subjectFilamentous hemagglutinin
dc.subjectPertussis toxin
dc.subjectVaccination
dc.subjectWhole blood
dc.titleA novel whole blood assay to quantify the release of T cell associated cytokines in response to Bordetella pertussis antigenseng
dc.typecontribution to journal
dspace.entity.typePublication
oaire.citation.startPage113758
oaire.citation.titleJournal of Immunological Methods
oaire.citation.volume534
oaire.versionhttp://purl.org/coar/version/c_970fb48d4fbd8a85

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