Enterovirus D68 infection induces IL-17–dependent neutrophilic airway inflammation and hyperresponsiveness
Charu Rajput, … , Emily T. Martin, Marc B. Hershenson
Charu Rajput, … , Emily T. Martin, Marc B. Hershenson
Published August 23, 2018
Citation Information: JCI Insight. 2018;3(16):e121882. https://doi.org/10.1172/jci.insight.121882.
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Research Article Infectious disease Pulmonology

Enterovirus D68 infection induces IL-17–dependent neutrophilic airway inflammation and hyperresponsiveness

Abstract

Enterovirus D68 (EV-D68) shares biologic features with rhinovirus (RV). In 2014, a nationwide outbreak of EV-D68 was associated with severe asthma-like symptoms. We sought to develop a mouse model of EV-D68 infection and determine the mechanisms underlying airway disease. BALB/c mice were inoculated intranasally with EV-D68 (2014 isolate), RV-A1B, or sham, alone or in combination with anti–IL-17A or house dust mite (HDM) treatment. Like RV-A1B, lung EV-D68 viral RNA peaked 12 hours after infection. EV-D68 induced airway inflammation, expression of cytokines (TNF-α, IL-6, IL-12b, IL-17A, CXCL1, CXCL2, CXCL10, and CCL2), and airway hyperresponsiveness, which were suppressed by anti–IL-17A antibody. Neutrophilic inflammation and airway responsiveness were significantly higher after EV-D68 compared with RV-A1B infection. Flow cytometry showed increased lineage–, NKp46–, RORγt+ IL-17+ILC3s and γδ T cells in the lungs of EV-D68–treated mice compared with those in RV-treated mice. EV-D68 infection of HDM-exposed mice induced additive or synergistic increases in BAL neutrophils and eosinophils and expression of IL-17, CCL11, IL-5, and Muc5AC. Finally, patients from the 2014 epidemic period with EV-D68 showed significantly higher nasopharyngeal IL-17 mRNA levels compared with patients with RV-A infection. EV-D68 infection induces IL-17–dependent airway inflammation and hyperresponsiveness, which is greater than that generated by RV-A1B, consistent with the clinical picture of severe asthma-like symptoms.

Authors

Charu Rajput, Mingyuan Han, J. Kelley Bentley, Jing Lei, Tomoko Ishikawa, Qian Wu, Joanna L. Hinde, Amy P. Callear, Terri L. Stillwell, William T. Jackson, Emily T. Martin, Marc B. Hershenson

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Figure 4

Flow cytometric analysis of IL-17–producing lung cells.

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Flow cytometric analysis of IL-17–producing lung cells. Female 8- to 10-...
Female 8- to 10-week-old BALB/c mice were treated with sham, 5 × 106 ePFU of EV-D68, or 5 × 106 PFU of RV-A1B. Forty-eight hours later, lungs were harvested, digested with collagenase, and stained with lineage antibody cocktail, anti-NKp46, anti-TCRγδ, anti-RORγt, anti–IL-17, and Pacific Blue (for dead cells). Cells were washed, fixed, and processed for flow cytometry. (A) Flow cytometry of live IL-17+ cells in sham, EV-D68, and RV-A1B groups. Group mean data are also shown. (B) Dot blots of lineage+ and lineage– live cells for the 3 conditions. (C) NKp46 and RORγt staining of lineage– cells. Group mean data for NKp46–, RORγt+, and NKp46 and RORγt double-positive cells are also shown. NKp46– and RORγt+ double-positive cells are gated for IL-17 staining. (D) TCR γδ and RORγt staining of lineage+ cells. Group mean data are also shown. TCR γδ and RORγt double-positive cells are gated for IL-17 staining. Data are shown as mean ± SEM of 6–7 mice/group from 2 separate experiments; *P < 0.05 by 1-way ANOVA, compared with sham; †P < 0.05 by 1-way ANOVA, compared with RV-1B.