Human Dectin-1 deficiency impairs macrophage-mediated defense against phaeohyphomycosis
Rebecca A. Drummond, … , Steven M. Holland, Michail S. Lionakis
Rebecca A. Drummond, … , Steven M. Holland, Michail S. Lionakis
Published November 15, 2022
Citation Information: J Clin Invest. 2022;132(22):e159348. https://doi.org/10.1172/JCI159348.
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Research Article Immunology Infectious disease

Human Dectin-1 deficiency impairs macrophage-mediated defense against phaeohyphomycosis

Abstract

Subcutaneous phaeohyphomycosis typically affects immunocompetent individuals following traumatic inoculation. Severe or disseminated infection can occur in CARD9 deficiency or after transplantation, but the mechanisms protecting against phaeohyphomycosis remain unclear. We evaluated a patient with progressive, refractory Corynespora cassiicola phaeohyphomycosis and found that he carried biallelic deleterious mutations in CLEC7A encoding the CARD9-coupled, β-glucan–binding receptor, Dectin-1. The patient’s PBMCs failed to produce TNF-α and IL-1β in response to β-glucan and/or C. cassiicola. To confirm the cellular and molecular requirements for immunity against C. cassiicola, we developed a mouse model of this infection. Mouse macrophages required Dectin-1 and CARD9 for IL-1β and TNF-α production, which enhanced fungal killing in an interdependent manner. Deficiency of either Dectin-1 or CARD9 was associated with more severe fungal disease, recapitulating the human observation. Because these data implicated impaired Dectin-1 responses in susceptibility to phaeohyphomycosis, we evaluated 17 additional unrelated patients with severe forms of the infection. We found that 12 out of 17 carried deleterious CLEC7A mutations associated with an altered Dectin-1 extracellular C-terminal domain and impaired Dectin-1–dependent cytokine production. Thus, we show that Dectin-1 and CARD9 promote protective TNF-α– and IL-1β–mediated macrophage defense against C. cassiicola. More broadly, we demonstrate that human Dectin-1 deficiency may contribute to susceptibility to severe phaeohyphomycosis by certain dematiaceous fungi.

Authors

Rebecca A. Drummond, Jigar V. Desai, Amy P. Hsu, Vasileios Oikonomou, Donald C. Vinh, Joshua A. Acklin, Michael S. Abers, Magdalena A. Walkiewicz, Sarah L. Anzick, Muthulekha Swamydas, Simon Vautier, Mukil Natarajan, Andrew J. Oler, Daisuke Yamanaka, Katrin D. Mayer-Barber, Yoichiro Iwakura, David Bianchi, Brian Driscoll, Ken Hauck, Ahnika Kline, Nicholas S.P. Viall, Christa S. Zerbe, Elise M.N. Ferré, Monica M. Schmitt, Tom DiMaggio, Stefania Pittaluga, John A. Butman, Adrian M. Zelazny, Yvonne R. Shea, Cesar A. Arias, Cameron Ashbaugh, Maryam Mahmood, Zelalem Temesgen, Alexander G. Theofiles, Masayuki Nigo, Varsha Moudgal, Karen C. Bloch, Sean G. Kelly, M. Suzanne Whitworth, Ganesh Rao, Cindy J. Whitener, Neema Mafi, Juan Gea-Banacloche, Lawrence C. Kenyon, William R. Miller, Katia Boggian, Andrea Gilbert, Matthew Sincock, Alexandra F. Freeman, John E. Bennett, Rodrigo Hasbun, Constantinos M. Mikelis, Kyung J. Kwon-Chung, Yasmine Belkaid, Gordon D. Brown, Jean K. Lim, Douglas B. Kuhns, Steven M. Holland, Michail S. Lionakis

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Figure 1

A Dectin-1–deficient patient with severe Corynespora cassiicola phaeohyphomycosis.

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A Dectin-1–deficient patient with severe Corynespora cassiicola phaeohyp...
(A) Photographs of the index patient at presentation at the NIH in 2004 (left) and following antifungal treatment and secondary prophylaxis in 2018 (right). (B) Grocott’s methenamine silver–stained (GMS-stained, left) and hematoxylin and eosin–stained (H&E-stained, right) section of soft tissue biopsy demonstrating C. cassiicola being engulfed but not destroyed in macrophages (black arrowheads) within granulomas (×100 magnification shown in Supplemental Figure 1). Scale bars: 20 μm. (C) Chromatograms from CLEC7A sequencing on healthy control and our patient, over the site of mutation in each allele. (D) Representative FACS histograms showing Dectin-1 surface expression in our patient and 2 healthy control patients. Histograms were gated on CD14+ monocytes isolated from peripheral blood. (E) Representative protein immunoblot images of Dectin-1 expression in PBMCs from our patient and a healthy volunteer (HV). β-Actin was used as loading control. (F) TNF-α production by PBMCs after 48 hours of stimulation with either purified particulate β-glucan or α-mannan. Each data point represents an individual well; at least 2 separate blood draws were analyzed in the Dectin-1–deficient patient (each tested in 2–3 technical replicates) and compared to 2 different healthy controls (each tested in 2–3 technical replicates). A single blood draw from each of the CARD9-deficient patients was analyzed in 3–5 technical replicates. Data in panel F were analyzed by 2-way ANOVA with Bonferroni’s correction. ***P < 0.005, ****P < 0.0001. NS, not significant. (G) Photograph of a previously reported CARD9-deficient patient (CARD9.02) (21) at presentation at the NIH (age 12). (H) Volume rendering of computed tomography data of patient CARD9.02 at age 12 emphasizing bone, which reveals erosions of the frontal bone (black arrows) and loss of maxillofacial structures (white arrow), including the hard palate, resulting in a common oronasal cavity. (I) Parasagittal T1-weighted magnetic resonance imaging of patient CARD9.02 at age 12 obtained following i.v. gadolinium-based contrast agent administration, which reveals epidural abscess with adjacent cerebritis (black arrow) and tissue loss (white arrow) resulting in a common cavity encompassing the nasopharynx, oropharynx, nasal cavity, oral cavity, and portions of the paranasal sinuses. (J) GMS- (upper) and H&E-stained (lower) section of soft tissue biopsy demonstrating granulomatous inflammation with C. cassiicola engulfed within macrophages (accompanying images from brain biopsy shown in Supplemental Figure 4). Both images are from consecutive cuts of the same biopsy sample. Scale bars: 20 μm.