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Development of a simple and robust whole blood assay with dual co-stimulation to quantify the release of T-cellular signature cytokines in response to Aspergillus fumigatus antigens

Please always quote using this URN: urn:nbn:de:bvb:20-opus-241025
  • Deeper understanding of mold-induced cytokine signatures could promote advances in the diagnosis and treatment of invasive mycoses and mold-associated hypersensitivity syndromes. Currently, most T-cellular immunoassays in medical mycology require the isolation of mononuclear cells and have limited robustness and practicability, hampering their broader applicability in clinical practice. Therefore, we developed a simple, cost-efficient whole blood (WB) assay with dual α-CD28 and α-CD49d co-stimulation to quantify cytokine secretion in responseDeeper understanding of mold-induced cytokine signatures could promote advances in the diagnosis and treatment of invasive mycoses and mold-associated hypersensitivity syndromes. Currently, most T-cellular immunoassays in medical mycology require the isolation of mononuclear cells and have limited robustness and practicability, hampering their broader applicability in clinical practice. Therefore, we developed a simple, cost-efficient whole blood (WB) assay with dual α-CD28 and α-CD49d co-stimulation to quantify cytokine secretion in response to Aspergillus fumigatus antigens. Dual co-stimulation strongly enhanced A. fumigatus-induced release of T-cellular signature cytokines detectable by enzyme-linked immunosorbent assay (ELISA) or a multiplex cytokine assay. Furthermore, T-cell-dependent activation and cytokine response of innate immune cells was captured by the assay. The protocol consistently showed little technical variation and high robustness to pre-analytic delays of up to 8 h. Stimulation with an A. fumigatus lysate elicited at least 7-fold greater median concentrations of key T-helper cell signature cytokines, including IL-17 and the type 2 T-helper cell cytokines IL-4 and IL-5 in WB samples from patients with Aspergillus-associated lung pathologies versus patients with non-mold-related lung diseases, suggesting high discriminatory power of the assay. These results position WB-ELISA with dual co-stimulation as a simple, accurate, and robust immunoassay for translational applications, encouraging further evaluation as a platform to monitor host immunity to opportunistic pathogens.show moreshow less

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Metadaten
Author: Chris D. Lauruschkat, Lukas Page, P. Lewis White, Sonja Etter, Helen E. Davies, Jamie Duckers, Frank Ebel, Elisabeth Schnack, Matthijs Backx, Mariola Dragan, Nicolas Schlegel, Olaf Kniemeyer, Axel A. Brakhage, Hermann Einsele, Juergen Loeffler, Sebastian Wurster
URN:urn:nbn:de:bvb:20-opus-241025
Document Type:Journal article
Faculties:Medizinische Fakultät / Klinik und Poliklinik für Allgemein-, Viszeral-, Gefäß- und Kinderchirurgie (Chirurgische Klinik I)
Medizinische Fakultät / Medizinische Klinik und Poliklinik II
Language:English
Parent Title (English):Journal of Fungi
ISSN:2309-608X
Year of Completion:2021
Volume:7
Issue:6
Article Number:462
Source:Journal of Fungi (2021) 7:6, 462. https://doi.org/10.3390/jof7060462
DOI:https://doi.org/10.3390/jof7060462
Dewey Decimal Classification:6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit
Tag:Aspergillus; adaptive immunity; biomarker; cytokines; immunoassay; inflammation
Release Date:2022/08/03
Date of first Publication:2021/06/08
Licence (German):License LogoCC BY: Creative-Commons-Lizenz: Namensnennung 4.0 International