A1 Refereed original research article in a scientific journal
A novel whole blood assay to quantify the release of T cell associated cytokines in response to Bordetella pertussis antigens
Authors: Pinto, Marta Valente; Barkoff, Alex-Mikael; Bibi, Sagida; Knuutila, Aapo; Teräsjärvi, Johanna; Clutterbuck, Elizabeth; Gimenez-Fourage, Sophie; Pagnon, Anke; van Gaans-van den Brink, Jacqueline A.M.; Corbiere, Veronique; De Montfort, Aymeric; Saso, Anja; Jobe, Haddijatou; Roetynck, Sophie; Kampmann, Beate; Simonetti, Elles; Diavatopoulos, Dimitri; Lambert, Eleonora E.; Mertsola, Jussi; Blanc, Pascal; van Els, Cécile A. C. M.; Dominic Kelly, Dominic; He, Qiushui; The PERISCOPE Consortium
Publisher: Elsevier BV
Publishing place: AMSTERDAM
Publication year: 2024
Journal: Journal of Immunological Methods
Journal name in source: Journal of Immunological Methods
Journal acronym: J IMMUNOL METHODS
Article number: 113758
Volume: 534
Number of pages: 7
ISSN: 0022-1759
eISSN: 1872-7905
DOI: https://doi.org/10.1016/j.jim.2024.113758
Web address : https://doi.org/10.1016/j.jim.2024.113758
Self-archived copy’s web address: https://research.utu.fi/converis/portal/detail/Publication/458904586
Background: 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
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Funding information in the publication:
This research was funded by Innovative Medicines Initiative, grant number 115910 (European-Union), under PeriScope-project, the Tampere Tuberculosis Foundation, grant number 26006205 (Finland), and the Sigrid Juselius Foundation, grant number 240045 (Finland).
