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Innate lymphoid cells contribute to allergic airway disease exacerbation by obesity  Laetitia Everaere, PhD, Saliha Ait-Yahia, PhD, Olivier Molendi-Coste,

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Presentation on theme: "Innate lymphoid cells contribute to allergic airway disease exacerbation by obesity  Laetitia Everaere, PhD, Saliha Ait-Yahia, PhD, Olivier Molendi-Coste,"— Presentation transcript:

1 Innate lymphoid cells contribute to allergic airway disease exacerbation by obesity 
Laetitia Everaere, PhD, Saliha Ait-Yahia, PhD, Olivier Molendi-Coste, PhD, Han Vorng, BSc, Sandrine Quemener, MSc, Pauline LeVu, MSc, Sebastien Fleury, BSc, Emmanuel Bouchaert, BSc, Ying Fan, PhD, Catherine Duez, PhD, Patricia de Nadai, PhD, Bart Staels, DPhSc, David Dombrowicz, PhD, Anne Tsicopoulos, MD  Journal of Allergy and Clinical Immunology  Volume 138, Issue 5, Pages e11 (November 2016) DOI: /j.jaci Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 AAI exacerbation by obesity. A, AHR. B, BAL cytology. C, Representative PAS-, May-Grunwald Giemsa (MGG)–, and anti–major basic protein (MBP)–stained lung sections. D, Total and antigen-specific IgE concentrations. E, Lung cytokine and chemokine mRNA relative (Rel.) expression. Data are from 2 independent experiments (n = 7-14 mice per group) expressed as means ± SEMs. *P < .05, **P < .01, and ***P < .001, HDM versus PBS. #P < .05, ##P < .01, and ###P < .001, HFD versus LFD. Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Lung ILC2s, ILC3s, and CD4+ T cells in the setting of AAI exacerbation caused by obesity. A, Lung cell numbers and their cytokine expression. B, Lung protein or mRNA relative (Rel.) expression of ILC-activating cytokines or transcription factors. Data are from 2 independent experiments (n = 5-10 per group) expressed as means ± SEMs. *P < .05, **P < .01, and ***P < .001, HDM versus PBS. #P < .05, ##P < .01, and ###P < .001, HFD versus LFD. Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 VAT mRNA expression and ILC2s, ILC3s, and CD4+ T cells in the setting of AAI exacerbation caused by obesity. A, VAT mRNA relative (Rel.) expression of cytokines and transcription factors. Bars represent medians ± interquartile ranges (n = 8-19 per group). *P < .05, HDM versus PBS. #P < .05, ##P < .01, and ###P < .001, HFD versus LFD. B, VAT ILC2s, ILC3s, and CD4+ T cells as percentage of CD45+ cells (data from 2 pools of 7-8 mice). Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Effects of ILC depletion in the setting of AAI exacerbation caused by obesity. A, AHR. B, BAL cytology. C, Representative anti–major basic protein (MBP)–stained lung sections. D, Total IgE concentrations. E, Cytokine expression in lung CD4+ T cells. Data are expressed as means ± SEMs (n=4-11 per group). *P < .05, **P < .01, and ***P < .001, HDM versus PBS. #P < .05, ##P < .01, and ###P < .001, HFD versus LFD. ¤¤P < .01 and ¤¤¤P < .001, anti-CD90.2 versus isotype treatment. Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig E1 Experimental design. A, Basic protocol. B, Depletion protocol. Representative flow cytometric dot plots of ILC2 and NCR− ILC3 populations after anti-CD90.2 or isotype control antibody administration. ICOS, Inducible costimulator; i.n., intranasal; SSC, side scatter. Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig E2 Identification of lymphoid populations in the lung. Gating strategy for the different ILC and CD4 T-cell subsets included successively forward- and side-scatter gating, exclusion of multiplets, selection of living cells with a viability marker, and selection of subsets based on cell-surface markers. Example from a representative HFD-fed allergic mouse. FSC, Forward scatter; SSC, side scatter. Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig E3 Histologic analysis of the lung. A and B, Quantification of mucus (Fig E3, A) and eosinophil staining (Fig E3, B). Data are from 3 independent experiments and at least 3 to 5 different samples for each group and expressed as means ± SEMs. **P < .01 and ***P < .001, HDM versus PBS. #P < .05 and ###P < .001, HFD versus LFD. Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig E4 Antibody production and lung cytokine and chemokine expression. A, Antigen-specific IgG1 concentrations. B-F, Chemokine levels and cytokine mRNA relative (Rel.) expression were evaluated in sera and whole-lung lysates by using ELISA and quantitative RT-PCR, respectively. Data are from 2 independent experiments (n = 7-10 mice per group) and expressed as means ± SEMs. *P < .05 and ***P < .001, HDM versus PBS. ##P < .01 and ###P < .001, HFD versus LFD. Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Fig E5 A and B, Intracellular expression of IL-5 (Fig E5, A) and IL-4 (Fig E5, B) by lung ILC2s. Mean numbers and representative flow cytometric dot plots of IL-5+ and IL-4+ ILC2s in the lungs of PBS- and HDM-challenged LFD- and HFD-fed mice. Percentages values are indicated after subtraction of the control isotype. Data are expressed as means ± SEMs (n = 3-8 mice per group). *P < .05, HDM versus PBS. #P < .05 and ###P < .001, HFD versus LFD. Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

11 Fig E6 Pulmonary ILC-related cytokines or transcription factors. A and B, mRNA relative (Rel.) expression of cytokines or transcription factors associated with ILC2s (Fig E6, A) and ILC3s (Fig E6, B) from whole lungs. Data are from 2 independent experiments (n = 7-10 mice per group) and expressed as means ± SEMs. Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

12 Fig E7 AAI induction after lymphocyte reconstitution in Rag2-deficient mice. A, BAL cytology. B, Total IgE serum concentration. C, Lung ILC2 numbers and their cytokine expression. D, CD4+ T-cell numbers and their cytokine expression. Data are expressed as means ± SEMs (n = 7-8 per group). *P < .05, **P < .01, and ***P < .001, HDM versus PBS. ##P < .01 and ###P < .001, HFD versus LFD. Journal of Allergy and Clinical Immunology  , e11DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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