An IFN/STAT1/CYBB axis defines protective plasmacytoid DC–neutrophil crosstalk in Aspergillus fumigatus–infected mice
Yahui Guo, Mariano A. Aufiero, Kathleen A.M. Mills, Simon A. Grassmann, Hyunu Kim, Mergim Gjonbalaj, Paul Zumbo, Audrey Billips, Katrina B. Mar, Yao Yu, Laura C. Echeverri Tirado, Lena Heung, Amariliz Rivera, Doron Betel, Joseph C. Sun, Tobias M. Hohl
Yahui Guo, Mariano A. Aufiero, Kathleen A.M. Mills, Simon A. Grassmann, Hyunu Kim, Mergim Gjonbalaj, Paul Zumbo, Audrey Billips, Katrina B. Mar, Yao Yu, Laura C. Echeverri Tirado, Lena Heung, Amariliz Rivera, Doron Betel, Joseph C. Sun, Tobias M. Hohl
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Research Article Immunology Infectious disease

An IFN/STAT1/CYBB axis defines protective plasmacytoid DC–neutrophil crosstalk in Aspergillus fumigatus–infected mice

Abstract

Aspergillus fumigatus is the most common cause of invasive aspergillosis (IA), a devastating infection in immunocompromised patients. Plasmacytoid DCs (pDCs) regulate host defense against IA by enhancing neutrophil antifungal properties in the lung. Here, we define the pDC activation trajectory during A. fumigatus infection and the molecular events that underlie the protective pDC–neutrophil crosstalk. Fungus-induced pDC activation began after bone marrow egress and resulted in pDC-dependent regulation of lung type I and type III IFN levels. These pDC-derived products acted on type I and type III IFN receptor–expressing neutrophils and controlled neutrophil fungicidal activity and ROS production via STAT1 signaling in a cell-intrinsic manner. Mechanistically, neutrophil STAT1 signaling regulated transcription and expression of Cybb, which encodes one of 5 NADPH oxidase subunits. Thus, the results indicate that pDCs regulate neutrophil-dependent immunity against inhaled molds by controlling local expression of a subunit required for NADPH oxidase assembly and activity in the lung.

Authors

Yahui Guo, Mariano A. Aufiero, Kathleen A.M. Mills, Simon A. Grassmann, Hyunu Kim, Mergim Gjonbalaj, Paul Zumbo, Audrey Billips, Katrina B. Mar, Yao Yu, Laura C. Echeverri Tirado, Lena Heung, Amariliz Rivera, Doron Betel, Joseph C. Sun, Tobias M. Hohl

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Figure 4

pDCs regulate neutrophil STAT1–dependent antifungal activity.

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pDCs regulate neutrophil STAT1–dependent antifungal activity. (A) Scheme...
(A) Scheme to generate and to deplete pDCs or not in CD45.1+ BDCA2- DTRTg/+ Stat1+/+ and CD45.2+BDCA2–DTRTg/+ Stat1–/– mixed BM chimeric mice, resulting in 4 experimental groups (G1–G4). (B–D) Representative plots (B) display DsRed and AF633 fluorescence intensity in lung neutrophils from 4 experimental groups: G1, pDC+ Stat1+/+ (Stat1+/+ neutrophils from pDC-sufficient mice); G2, pDC– Stat1+/+ (Stat1+/+ neutrophils from pDC-depleted mice); G3, pDC+ Stat1–/– (Stat1–/– neutrophils from pDC-sufficient mice); G4, pDC– Stat1–/– (Stat1–/– neutrophils from pDC-depleted mice). The R1 gate indicates the frequency of neutrophils that contain live conidia; gate R2 indicates the frequency of neutrophils that contain killed conidia. (C and D) The plots show mean neutrophil (C) conidial uptake (R1 + R2) ± SEM and (D) conidial viability (R1/[R1 + R2]) ± SEM in indicated lung neutrophils isolated from the 4 groups. n = 12 per group. (E) Mean ± SEM ROS production in indicated lung neutrophils; n = 6 per group. B–D: Infection dose: 3 × 107 Af293 FLARE conidia via intratracheal route; analysis 72 hpi. (C and D) Data were pooled from 2 independent experiments and presented as mean ± SEM, (E) Data are representative of 2 experiments. Data points represent individual mice. Statistical analysis: Kruskal-Wallis test. See also Supplemental Figure 3.