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Article

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

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Department of Internal Medicine II, University Hospital of Wuerzburg, 97080 Wuerzburg, Germany
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Public Health Wales, Microbiology Cardiff, Wales CF14 4XW, UK
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Department of Respiratory Medicine, University Hospital of Wales, Llandough Site, Cardiff CF64 2XX, UK
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Cystic Fibrosis Unit, University Hospital of Wales, Llandough Site, Cardiff CF64 2XX, UK
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Institute for Infectious Diseases and Zoonoses, Ludwig-Maximilians-University of Munich, 80539 Munich, Germany
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Department of Surgery I, University Hospital of Wuerzburg, 97080 Wuerzburg, Germany
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Department of Molecular and Applied Microbiology, Leibniz Institute for Natural Product Research and Infection Biology–Hans-Knöll-Institute (HKI), 07745 Jena, Germany
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Department of Infectious Diseases, Infection Control and Employee Health, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA
*
Authors to whom correspondence should be addressed.
Academic Editor: George Deepe
J. Fungi 2021, 7(6), 462; https://doi.org/10.3390/jof7060462
Received: 20 April 2021 / Revised: 28 May 2021 / Accepted: 28 May 2021 / Published: 8 June 2021
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. View Full-Text
Keywords: immunoassay; biomarker; Aspergillus; cytokines; inflammation; adaptive immunity immunoassay; biomarker; Aspergillus; cytokines; inflammation; adaptive immunity
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MDPI and ACS Style

Lauruschkat, C.D.; Page, L.; White, P.L.; Etter, S.; Davies, H.E.; Duckers, J.; Ebel, F.; Schnack, E.; Backx, M.; Dragan, M.; Schlegel, N.; Kniemeyer, O.; Brakhage, A.A.; Einsele, H.; Loeffler, J.; Wurster, S. 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. J. Fungi 2021, 7, 462. https://doi.org/10.3390/jof7060462

AMA Style

Lauruschkat CD, Page L, White PL, Etter S, Davies HE, Duckers J, Ebel F, Schnack E, Backx M, Dragan M, Schlegel N, Kniemeyer O, Brakhage AA, Einsele H, Loeffler J, Wurster S. 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. Journal of Fungi. 2021; 7(6):462. https://doi.org/10.3390/jof7060462

Chicago/Turabian Style

Lauruschkat, Chris D., Lukas Page, P. L. 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, and Sebastian Wurster. 2021. "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" Journal of Fungi 7, no. 6: 462. https://doi.org/10.3390/jof7060462

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