TY  - JOUR
A1  - Lauruschkat, Chris D.
A1  - Page, Lukas
A1  - White, P. Lewis
A1  - Etter, Sonja
A1  - Davies, Helen E.
A1  - Duckers, Jamie
A1  - Ebel, Frank
A1  - Schnack, Elisabeth
A1  - Backx, Matthijs
A1  - Dragan, Mariola
A1  - Schlegel, Nicolas
A1  - Kniemeyer, Olaf
A1  - Brakhage, Axel A.
A1  - Einsele, Hermann
A1  - Loeffler, Juergen
A1  - Wurster, Sebastian
T1  - 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
T2  - Journal of Fungi
N2  - 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 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.
KW  - immunoassay
KW  - biomarker
KW  - Aspergillus
KW  - cytokines
KW  - inflammation
KW  - adaptive immunity
Y1  - 2021
UR  - https://opus.bibliothek.uni-wuerzburg.de/frontdoor/index/index/docId/24102
UR  - https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-241025
SN  - 2309-608X
VL  - 7
IS  - 6
ER  -