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Lung T-cell responses to nontypeable Haemophilus influenzae in patients with chronic obstructive pulmonary disease  Paul T. King, MD, PhD, Steven Lim,

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Presentation on theme: "Lung T-cell responses to nontypeable Haemophilus influenzae in patients with chronic obstructive pulmonary disease  Paul T. King, MD, PhD, Steven Lim,"— Presentation transcript:

1 Lung T-cell responses to nontypeable Haemophilus influenzae in patients with chronic obstructive pulmonary disease  Paul T. King, MD, PhD, Steven Lim, BSc, Adrian Pick, MD, James Ngui, BSc, Zdenka Prodanovic, BSc, William Downey, MD, Cliff Choong, MD, Anthony Kelman, MD, Elizabeth Baranyai, BSc, Michelle Francis, BSc, Randall Moshinsky, MD, Philip G. Bardin, MD, PhD, Peter W. Holmes, MD, Stephen R. Holdsworth, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 131, Issue 5, Pages e14 (May 2013) DOI: /j.jaci Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Percentage of T cells expressing the 5 inflammatory mediators after stimulation with SEB in the non-COPD group (control) compared with the COPD group. A, TH cell (CD4) response. B, CTL (CD8) response. There was no significant difference in any mediator production between the control and COPD groups. Journal of Allergy and Clinical Immunology  , e14DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 T-cell response to NTHi in the non-COPD group (control) compared with the COPD group. The TH cell (CD4) response to NTHi stimulation (A) and the CTL cell (CD8) response to NTHi stimulation (B) in the control and COPD groups by production of 5 inflammatory mediators is shown. The percentage of cells producing TNF-α, IL-13, and IL-17 were significantly higher in the COPD group. Journal of Allergy and Clinical Immunology  , e14DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 The ratio of the percentage of TH cells producing IFN-γ compared with the percentage of TH cells producing IL-13 after stimulation with NTHi in the control (A) and COPD (B) groups is shown. There was no significant difference in control subjects, but IL-13 levels were significantly higher than IFN-γ levels in patients with COPD, which is consistent with a TH2 response to NTHi. Journal of Allergy and Clinical Immunology  , e14DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Response of subgroups to NTHi. To measure the effect of smoking, the control group was divided into subgroups of those without a significant smoking history (nonsmoker) and those with a significant smoking history (smoker) and compared with the COPD group. There was a progressive increase in cytokine production for TNF-α, IL-13, and IL-17 in TH cells (A) and CTLs (B). Significant differences were mainly between nonsmoking control subjects and patients with COPD. Journal of Allergy and Clinical Immunology  , e14DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig E1 Flow cytometric gating strategy. A, Total cell suspension obtained from the lung of a patient with COPD with gating on the cell population expressing CD45. B, CD45 cells expressing CD3, which are further analyzed into cells expressing (C) either CD4 or CD8. CD4 cells are then further analyzed for their production of IL-13 and IL-17 (after stimulation). Journal of Allergy and Clinical Immunology  , e14DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig E2 Example of cytokine staining in CD4 cells induced by SEB. The figure shows an example of cytokine staining in TH cells induced by SEB (same patient with COPD shown in Fig 1, B), demonstrating production of TNF-α, IL-13, IL-17, CD154, and IFN-γ in unstimulated cells (left column) and after stimulation with SEB (right column). Journal of Allergy and Clinical Immunology  , e14DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig E3 Example of cytokine staining induced in CD4 cells by NTHi. The figure shows an example of cytokine staining in TH cells induced by NTHi (same subject shown in Fig 1, B), demonstrating production of TNF-α, IL-13, IL-17, CD154, and IFN-γ in unstimulated cells (left column) and after stimulation with NTHi (right column). Journal of Allergy and Clinical Immunology  , e14DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig E4 Example of cytokine staining induced in CD8 cells by NTHi. The figure shows an example of cytokine staining in TH cells induced by NTHi (same subject shown in Fig 1, B), demonstrating production of TNF-α, IL-13, IL-17, CD154, and IFN-γ in unstimulated cells (left column) and after stimulation with NTHi (right column). Journal of Allergy and Clinical Immunology  , e14DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Fig E5 Bead array results for T-cell cytokines from supernatants induced by NTHi stimulation. Cytokine levels were compared between the non-COPD (control) and COPD groups for TNF-α (P = .16), IL-13 (P = .06), IL-17 (P = .01), and IFN-γ (P = .31). Journal of Allergy and Clinical Immunology  , e14DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

11 Fig E6 Bead array results for innate immune mediator production from supernatants induced by NTHi stimulation. Cytokine levels were compared between the non-COPD (control) and COPD groups for IL-1β (P = .65), IL-6 (P = .26), and IL-12 (P = .79). Journal of Allergy and Clinical Immunology  , e14DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

12 Fig E7 Bead array results for IL-10 production from supernatants induced by NTHi stimulation. Cytokine levels of IL-10 were compared between the non-COPD (control) and COPD groups (P = .90). Journal of Allergy and Clinical Immunology  , e14DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

13 Fig E8 The ratio of T cells (CD45+, CD3+) to the population of white cells that were CD45+ but CD3− (these CD45+CD3− cells would comprise most of the classical APCs) among the 3 groups (nonsmoking control subjects, smoking control subjects, and patients with COPD). ANOVA demonstrated no significant differences (P = .63). Journal of Allergy and Clinical Immunology  , e14DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

14 Fig E9 Bead array results for IL-2 (A) and IL-9 (B) production in supernatants induced by NTHi stimulation. Cytokine levels of IL-2 were similar between the non-COPD (control) and COPD groups (P = .29), as were IL-9 levels (P = .24). Journal of Allergy and Clinical Immunology  , e14DOI: ( /j.jaci ) Copyright © 2012 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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