TY  - JOUR
A1  - Tappe, Beeke
A1  - Lauruschkat, Chris D.
A1  - Strobel, Lea
A1  - Pantaleón García, Jezreel
A1  - Kurzai, Oliver
A1  - Rebhan, Silke
A1  - Kraus, Sabrina
A1  - Pfeuffer-Jovic, Elena
A1  - Bussemer, Lydia
A1  - Possler, Lotte
A1  - Held, Matthias
A1  - Hünniger, Kerstin
A1  - Kniemeyer, Olaf
A1  - Schäuble, Sascha
A1  - Brakhage, Axel A.
A1  - Panagiotou, Gianni
A1  - White, P. Lewis
A1  - Einsele, Hermann
A1  - Löffler, Jürgen
A1  - Wurster, Sebastian
T1  - COVID-19 patients share common, corticosteroid-independent features of impaired host immunity to pathogenic molds
JF  - Frontiers in Immunology
N2  - Patients suffering from coronavirus disease-2019 (COVID-19) are susceptible to deadly secondary fungal infections such as COVID-19-associated pulmonary aspergillosis and COVID-19-associated mucormycosis. Despite this clinical observation, direct experimental evidence for severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2)-driven alterations of antifungal immunity is scarce. Using an ex-vivo whole blood stimulation assay, we challenged blood from twelve COVID-19 patients with Aspergillus fumigatus and Rhizopus arrhizus antigens and studied the expression of activation, maturation, and exhaustion markers, as well as cytokine secretion. Compared to healthy controls, T-helper cells from COVID-19 patients displayed increased expression levels of the exhaustion marker PD-1 and weakened A. fumigatus- and R. arrhizus-induced activation. While baseline secretion of proinflammatory cytokines was massively elevated, whole blood from COVID-19 patients elicited diminished release of T-cellular (e.g., IFN-γ, IL-2) and innate immune cell-derived (e.g., CXCL9, CXCL10) cytokines in response to A. fumigatus and R. arrhizus antigens. Additionally, samples from COVID-19 patients showed deficient granulocyte activation by mold antigens and reduced fungal killing capacity of neutrophils. These features of weakened anti-mold immune responses were largely decoupled from COVID-19 severity, the time elapsed since diagnosis of COVID-19, and recent corticosteroid uptake, suggesting that impaired anti-mold defense is a common denominator of the underlying SARS-CoV-2 infection. Taken together, these results expand our understanding of the immune predisposition to post-viral mold infections and could inform future studies of immunotherapeutic strategies to prevent and treat fungal superinfections in COVID-19 patients.
KW  - COVID-19
KW  - immune impairment
KW  - T cells
KW  - granulocytes
KW  - Aspergillus
KW  - Rhizopus
Y1  - 2022
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-283558
SN  - 1664-3224
VL  - 13
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  - 
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  - Yu, Yidong
A1  - Wolf, Ann-Katrin
A1  - Thusek, Sina
A1  - Heinekamp, Thorsten
A1  - Bromley, Michael
A1  - Krappmann, Sven
A1  - Terpitz, Ulrich
A1  - Voigt, Kerstin
A1  - Brakhage, Axel A.
A1  - Beilhack, Andreas
T1  - Direct Visualization of Fungal Burden in Filamentous Fungus-Infected Silkworms
JF  - Journal of Fungi
N2  - Invasive fungal infections (IFIs) are difficult to diagnose and to treat and, despite several available antifungal drugs, cause high mortality rates. In the past decades, the incidence of IFIs has continuously increased. More recently, SARS-CoV-2-associated lethal IFIs have been reported worldwide in critically ill patients. Combating IFIs requires a more profound understanding of fungal pathogenicity to facilitate the development of novel antifungal strategies. Animal models are indispensable for studying fungal infections and to develop new antifungals. However, using mammalian animal models faces various hurdles including ethical issues and high costs, which makes large-scale infection experiments extremely challenging. To overcome these limitations, we optimized an invertebrate model and introduced a simple calcofluor white (CW) staining protocol to macroscopically and microscopically monitor disease progression in silkworms (Bombyx mori) infected with the human pathogenic filamentous fungi Aspergillus fumigatus and Lichtheimia corymbifera. This advanced silkworm A. fumigatus infection model could validate knockout mutants with either attenuated, strongly attenuated or unchanged virulence. Finally, CW staining allowed us to efficiently visualize antifungal treatment outcomes in infected silkworms. Conclusively, we here present a powerful animal model combined with a straightforward staining protocol to expedite large-scale in vivo research of fungal pathogenicity and to investigate novel antifungal candidates.
KW  - fungal infection model
KW  - calcofluor white staining
KW  - Aspergillus
KW  - Lichtheimia
KW  - silkworm
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-228855
SN  - 2309-608X
VL  - 7
IS  - 2
ER  - 
TY  - JOUR
A1  - Page, Lukas
A1  - Wallstabe, Julia
A1  - Lother, Jasmin
A1  - Bauser, Maximilian
A1  - Kniemeyer, Olaf
A1  - Strobel, Lea
A1  - Voltersen, Vera
A1  - Teutschbein, Janka
A1  - Hortschansky, Peter
A1  - Morton, Charles Oliver
A1  - Brakhage, Axel A.
A1  - Topp, Max
A1  - Einsele, Hermann
A1  - Wurster, Sebastian
A1  - Loeffler, Juergen
T1  - CcpA- and Shm2-Pulsed Myeloid Dendritic Cells Induce T-Cell Activation and Enhance the Neutrophilic Oxidative Burst Response to Aspergillus fumigatus
JF  - Frontiers in Immunology
N2  - Aspergillus fumigatus causes life-threatening opportunistic infections in immunocompromised patients. As therapeutic outcomes of invasive aspergillosis (IA) are often unsatisfactory, the development of targeted immunotherapy remains an important goal. Linking the innate and adaptive immune system, dendritic cells are pivotal in anti-Aspergillus defense and have generated interest as a potential immunotherapeutic approach in IA. While monocyte-derived dendritic cells (moDCs) require ex vivo differentiation, antigen-pulsed primary myeloid dendritic cells (mDCs) may present a more immediate platform for immunotherapy. To that end, we compared the response patterns and cellular interactions of human primary mDCs and moDCs pulsed with an A. fumigatus lysate and two A. fumigatus proteins (CcpA and Shm2) in a serum-free, GMP-compliant medium. CcpA and Shm2 triggered significant upregulation of maturation markers in mDCs and, to a lesser extent, moDCs. Furthermore, both A. fumigatus proteins elicited the release of an array of key pro-inflammatory cytokines including TNF-α, IL-1β, IL-6, IL-8, and CCL3 from both DC populations. Compared to moDCs, CcpA- and Shm2-pulsed mDCs exhibited greater expression of MHC class II antigens and stimulated stronger proliferation and IFN-γ secretion from autologous CD4\(^+\) and CD8\(^+\) T-cells. Moreover, supernatants of CcpA- and Shm2-pulsed mDCs significantly enhanced the oxidative burst in allogeneic neutrophils co-cultured with A. fumigatus germ tubes. Taken together, our in vitro data suggest that ex vivo CcpA- and Shm2-pulsed primary mDCs have the potential to be developed into an immunotherapeutic approach to tackle IA.
KW  - antigens
KW  - dendritic cells
KW  - cytokines
KW  - host defense
KW  - immunotherapy
KW  - Aspergillus
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-239493
SN  - 1664-3224
VL  - 12
ER  - 
TY  - JOUR
A1  - Schmidt, Stefanie
A1  - Ebner, Friederike
A1  - Rosen, Kerstin
A1  - Kniemeyer, Olaf
A1  - Brakhage, Axel A.
A1  - Löffler, Jürgen
A1  - Seif, Michelle
A1  - Springer, Jan
A1  - Schlosser, Josephine
A1  - Scharek‐Tedin, Lydia
A1  - Scheffold, Alexander
A1  - Bacher, Petra
A1  - Kühl, Anja A.
A1  - Rösler, Uwe
A1  - Hartmann, Susanne
T1  - The domestic pig as human‐relevant large animal model to study adaptive antifungal immune responses against airborne Aspergillus fumigatus
JF  - European Journal of Immunology
N2  - Pulmonary mucosal immune response is critical for preventing opportunistic Aspergillus fumigatus infections. Although fungus‐specific CD4\(^{+}\) T cells in blood are described to reflect the actual host–pathogen interaction status, little is known about Aspergillus‐specific pulmonary T‐cell responses. Here, we exploit the domestic pig as human‐relevant large animal model and introduce antigen‐specific T‐cell enrichment in pigs to address Aspergillus‐specific T cells in the lung compared to peripheral blood. In healthy, environmentally Aspergillus‐exposed pigs, the fungus‐specific T cells are detectable in blood in similar frequencies as observed in healthy humans and exhibit a Th1 phenotype. Exposing pigs to 10\(^{6}\) cfu/m\(^{3}\) conidia induces a long‐lasting accumulation of Aspergillus‐specific Th1 cells locally in the lung and also systemically. Temporary immunosuppression during Aspergillus‐exposure showed a drastic reduction in the lung‐infiltrating antifungal T‐cell responses more than 2 weeks after abrogation of the suppressive treatment. This was reflected in blood, but to a much lesser extent. In conclusion, by using the human‐relevant large animal model the pig, this study highlights that the blood clearly reflects the mucosal fungal‐specific T‐cell reactivity in environmentally exposed as well as experimentally exposed healthy pigs. But, immunosuppression significantly impacts the mucosal site in contrast to the initial systemic immune response.
KW  - fungal aerosolization
KW  - porcine large animal model
KW  - pulmonary immune response
KW  - T cells
KW  - Aspergillus fumigatus
Y1  - 2020
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-216085
VL  - 50
IS  - 11
SP  - 1712
EP  - 1728
ER  -