BTK drives neutrophil activation for sterilizing antifungal immunity
Jigar V. Desai, Marissa A. Zarakas, Andrew L. Wishart, Mark Roschewski, Mariano A. Aufiero, Agnes Donkò, Gustaf Wigerblad, Neta Shlezinger, Markus Plate, Matthew R. James, Jean K. Lim, Gulbu Uzel, Jenna R.E. Bergerson, Ivan Fuss, Robert A. Cramer, Luis M. Franco, Emily S. Clark, Wasif N. Khan, Daisuke Yamanaka, Georgios Chamilos, Jamel El-Benna, Mariana J. Kaplan, Louis M. Staudt, Thomas L. Leto, Steven M. Holland, Wyndham H. Wilson, Tobias M. Hohl, Michail S. Lionakis
Jigar V. Desai, Marissa A. Zarakas, Andrew L. Wishart, Mark Roschewski, Mariano A. Aufiero, Agnes Donkò, Gustaf Wigerblad, Neta Shlezinger, Markus Plate, Matthew R. James, Jean K. Lim, Gulbu Uzel, Jenna R.E. Bergerson, Ivan Fuss, Robert A. Cramer, Luis M. Franco, Emily S. Clark, Wasif N. Khan, Daisuke Yamanaka, Georgios Chamilos, Jamel El-Benna, Mariana J. Kaplan, Louis M. Staudt, Thomas L. Leto, Steven M. Holland, Wyndham H. Wilson, Tobias M. Hohl, Michail S. Lionakis
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Research Article Immunology Infectious disease

BTK drives neutrophil activation for sterilizing antifungal immunity

Abstract

We describe a previously unappreciated role for Bruton’s tyrosine kinase (BTK) in fungal immune surveillance against aspergillosis, an unforeseen complication of BTK inhibitors (BTKi) used for treating B cell lymphoid malignancies. We studied BTK-dependent fungal responses in neutrophils from diverse populations, including healthy donors, patients who were treated with BTKi, and X-linked agammaglobulinemia patients. Upon fungal exposure, BTK was activated in human neutrophils in a TLR2-, Dectin-1-, and FcγR-dependent manner, triggering the oxidative burst. BTK inhibition selectively impeded neutrophil-mediated damage to Aspergillus hyphae, primary granule release, and the fungus-induced oxidative burst by abrogating NADPH oxidase subunit p40phox and GTPase RAC2 activation. Moreover, neutrophil-specific Btk deletion in mice enhanced aspergillosis susceptibility by impairing neutrophil function, not recruitment or lifespan. Conversely, GM-CSF partially mitigated these deficits by enhancing p47phox activation. Our findings underline the crucial role of BTK signaling in neutrophils for antifungal immunity and provide a rationale for GM-CSF use to offset these deficits in patients who are susceptible.

Authors

Jigar V. Desai, Marissa A. Zarakas, Andrew L. Wishart, Mark Roschewski, Mariano A. Aufiero, Agnes Donkò, Gustaf Wigerblad, Neta Shlezinger, Markus Plate, Matthew R. James, Jean K. Lim, Gulbu Uzel, Jenna R.E. Bergerson, Ivan Fuss, Robert A. Cramer, Luis M. Franco, Emily S. Clark, Wasif N. Khan, Daisuke Yamanaka, Georgios Chamilos, Jamel El-Benna, Mariana J. Kaplan, Louis M. Staudt, Thomas L. Leto, Steven M. Holland, Wyndham H. Wilson, Tobias M. Hohl, Michail S. Lionakis

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Figure 1

Neutrophil-specific BTK confers protection during pulmonary aspergillosis.

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Neutrophil-specific BTK confers protection during pulmonary aspergillosi...
(A) Survival of WT and Btk–/– mice after infection with A. fumigatus (n = 10–14). (B and C) Representative micrographs of (B) H&E-stained and (C) Grocott’s methenamine silver–stained (GMS-stained) lung sections at day 4 after infection. Scale bars: 1 mm (upper panels, B and C), 250 μm (lower panel, B), 25 μm (lower panel, C) (n = 4). (D) Quantification of the proportion of germinating A. fumigatus conidia in GMS-stained lung sections. Each dot depicts an individual affected region of the lung; 4 such areas were randomly chosen per mouse (n = 4 mice) and germinated conidia were enumerated. (E) β-D-glucan levels in lung homogenates at steady state and day 2 after infection. Each dot depicts an individual mouse (n = 5–10). (F) Survival of ibrutinib- or vehicle-treated WT and Rag2–/– mice after infection with A. fumigatus (n = 11–21). (G–I) Survival of the indicated mice after infection with A. fumigatus (G, n = 14–15, H, n = 18–28; I, n = 33–34). Box and whisker plots depict values ranging from minimum to maximum (D and E). *P<0.05, **P<0.01, ****P<0.0001, determined using log-rank test (A and F–I), 2-sided Mann-Whitney U test (D and E), or 2-sided unpaired t test (E).