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
JF  - 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
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-241025
SN  - 2309-608X
VL  - 7
IS  - 6
ER  - 
TY  - JOUR
A1  - Lauruschkat, Chris D.
A1  - Etter, Sonja
A1  - Schnack, Elisabeth
A1  - Ebel, Frank
A1  - Schäuble, Sascha
A1  - Page, Lukas
A1  - Rümens, Dana
A1  - Dragan, Mariola
A1  - Schlegel, Nicolas
A1  - Panagiotou, Gianni
A1  - Kniemeyer, Olaf
A1  - Brakhage, Axel A.
A1  - Einsele, Hermann
A1  - Wurster, Sebastian
A1  - Loeffler, Juergen
T1  - Chronic occupational mold exposure drives expansion of Aspergillus-reactive type 1 and type 2 T-helper cell responses
JF  - Journal of Fungi
N2  - Occupational mold exposure can lead to Aspergillus-associated allergic diseases including asthma and hypersensitivity pneumonitis. Elevated IL-17 levels or disbalanced T-helper (Th) cell expansion were previously linked to Aspergillus-associated allergic diseases, whereas alterations to the Th cell repertoire in healthy occupationally exposed subjects are scarcely studied. Therefore, we employed functional immunoassays to compare Th cell responses to A. fumigatus antigens in organic farmers, a cohort frequently exposed to environmental molds, and non-occupationally exposed controls. Organic farmers harbored significantly higher A. fumigatus-specific Th-cell frequencies than controls, with comparable expansion of Th1- and Th2-cell frequencies but only slightly elevated Th17-cell frequencies. Accordingly, Aspergillus antigen-induced Th1 and Th2 cytokine levels were strongly elevated, whereas induction of IL-17A was minimal. Additionally, increased levels of some innate immune cell-derived cytokines were found in samples from organic farmers. Antigen-induced cytokine release combined with Aspergillus-specific Th-cell frequencies resulted in high classification accuracy between organic farmers and controls. Aspf22, CatB, and CipC elicited the strongest differences in Th1 and Th2 responses between the two cohorts, suggesting these antigens as potential candidates for future bio-effect monitoring approaches. Overall, we found that occupationally exposed agricultural workers display a largely balanced co-expansion of Th1 and Th2 immunity with only minor changes in Th17 responses.
KW  - mold exposure
KW  - immunoassay
KW  - biomarker
KW  - Aspergillus
KW  - cytokines
KW  - inflammation
KW  - adaptive immunity
KW  - hypersensitivity
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-245202
SN  - 2309-608X
VL  - 7
IS  - 9
ER  -