Allergens abrogate antiinflammatory DNA effects and unmask macrophage-driven neutrophilic asthma via ILC2/STING/TNF-α signaling
Anand Sripada, Divya Verma, Rangati Varma, Kapil Sirohi, Carolyn Kwiat, Mohini Pathria, Mukesh Verma, Anita Sahu, Vamsi Guntur, Laurie Manka, Brian Vestal, Camille Moore, Richard J. Martin, Magdalena M. Gorska, John Cambier, Andrew Getahun, Rafeul Alam
Anand Sripada, Divya Verma, Rangati Varma, Kapil Sirohi, Carolyn Kwiat, Mohini Pathria, Mukesh Verma, Anita Sahu, Vamsi Guntur, Laurie Manka, Brian Vestal, Camille Moore, Richard J. Martin, Magdalena M. Gorska, John Cambier, Andrew Getahun, Rafeul Alam
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Research Article Immunology Inflammation Pulmonology

Allergens abrogate antiinflammatory DNA effects and unmask macrophage-driven neutrophilic asthma via ILC2/STING/TNF-α signaling

Abstract

The mechanisms of neutrophilic and mixed neutrophilic-eosinophilic asthma are poorly understood. We found that extracellular DNA and nucleosomes (Nucs) were elevated in the airways of patients with neutrophilic-eosinophilic asthma and correlated with bronchoalveolar lavage neutrophils. Bronchial tissue from neutrophilic-eosinophilic asthma had more DNA sensor–positive cells. Intranasally administered DNA did not induce airway hyperreactivity (AHR) or any pathology but induced AHR and neutrophilic-eosinophilic inflammation when coadministered with the allergen Alternaria (Alt). Nuc alone induced antiinflammatory/defensive genes, whereas the Nuc-Alt combination increased levels of TNF-α and innate cytokines. The Alt-Nuc phenotype was abolished in Cgas–/–, ALR–/–, Sting–/–, LysMCre:Stingfl/fl, IL7RCre:Rorαfl/fl, and Tnfr2–/– mice. Alt, unexpectedly, played an essential role in the Nuc-induced phenotype. It abrogated Nuc induction of antiinflammatory genes, facilitated Nuc uptake, induced type 2 innate lymphoid cells, which, in the presence of Nuc, produced high levels of TNF-α, and promoted neutrophilic infiltration. We established a paradigm whereby allergens inhibit the antiinflammatory effects of DNA/Nuc and facilitate STING-TNF-α–driven neutrophilic-eosinophilic inflammation in asthma.

Authors

Anand Sripada, Divya Verma, Rangati Varma, Kapil Sirohi, Carolyn Kwiat, Mohini Pathria, Mukesh Verma, Anita Sahu, Vamsi Guntur, Laurie Manka, Brian Vestal, Camille Moore, Richard J. Martin, Magdalena M. Gorska, John Cambier, Andrew Getahun, Rafeul Alam

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Figure 7

Effect of germline deletion of various DNA sensors and ILC2s on Alt-Nuc and Alt-induced asthma.

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Effect of germline deletion of various DNA sensors and ILC2s on Alt-Nuc ...
(A) AHR after Alt-Nuc, Alt, and Sal exposure in WT and Cgas–/– mice (n = 5) as measured by flexiVent. RRS, respiratory system resistance. εWT-Sal versus WT-Alt-Nuc, P < 0.0001; #WT-Sal versus WT-Alt, P < 0.0001; §WT-Sal versus Cgas–/–-Alt, P < 0.0001; *WT-Sal versus Cgas–/–-Alt-Nuc, P = 0.001; ΨWT-Alt-Nuc versus Cgas–/–-Alt-Nuc, P < 0.0001; ΦWT-Alt versus WT-Alt-Nuc, P < 0.0001. (B–E) Quantification of eosinophils, neutrophils, lung inflammation, and mucus production in WT and Cgas–/– mice treated with Alt or Alt-Nuc (n = 5). (Groups are color-coded as in A). BM, basement membrane. (F) AHR after Alt-Nuc and Sal exposure in WT and ALR–/– mice (n = 5). *WT-Sal versus WT-Alt-Nuc, P < 0.0001; δWT-Sal versus ALR–/–-Alt-Nuc, P = 0.0003; ΨWT-Alt-Nuc versus ALR–/–-Alt-Nuc, P < 0.0001. (G–J) Quantification of eosinophils, neutrophils, lung inflammation, and mucus production in WT and ALR–/– mice treated with Alt-Nuc (n = 5). (Groups are color-coded as in F). (K) AHR after Alt-Nuc, Alt and Sal exposure in Stingfl/fl and LysMCre:Stingfl/fl mice (n = 5). εStingfl/fl-Sal versus Stingfl/fl-Alt-Nuc, P < 0.0001; #Stingfl/fl-Sal versus Stingfl/fl -Alt, P < 0.0001; *Stingfl/fl-Sal versus LysMCre:Stingfl/fl-Alt, P < 0.0001; ΨStingfl/fl-Alt-Nuc versus LysMCre:Stingfl/fl-Alt-Nuc, P<0.0001; §LysMCre:Stingfl/fl-Alt versus LysMCre:Stingfl/fl-Alt-Nuc, P = 0.02. (L–O) Quantification of eosinophils, neutrophils, lung inflammation, and mucus production in Stingfl/fl and LysMCre:Stingfl/fl mice treated with Alt-Nuc, Alt, or Sal (n = 5). (Groups are color-coded as in K). (P) AHR after Alt-Nuc, Alt and Sal exposure in RORαfl/fl and IL7RCre:RORαfl/fl mice (n = 5). δRORαfl/fl-Sal versus RORαfl/fl-Alt-Nuc, P < 0.0001; #RORαfl/fl-Sal versus RORαfl/fl-Alt, P < 0.0001; §RORαfl/fl-Sal versus IL7RCre:RORαfl/fl-Alt, P = 0.007; ΨRORαfl/fl-Alt-Nuc versus IL7RCre:RORαfl/fl-Alt-Nuc, P < 0.0001; *RORαfl/fl-Alt versus RORαfl/fl-Alt-Nuc, P < 0.0001; ΦRORαfl/fl-Alt versus IL7RCre:RORαfl/fl-Alt, P = 0.01. (Q–T) Quantification of eosinophils, neutrophils, lung inflammation, and mucus production in RORαfl/fl and IL7RCre:RORαfl/fl mice (n = 5) treated with Alt-Nuc, Alt, and Sal (groups are color-coded as in P). Two-way ANOVA with Tukey’s multiple comparisons test was used to determine the statistical significance between groups. Data are presented as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 ****P < 0.0001.