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Interferon response and respiratory virus control are preserved in bronchial epithelial cells in asthma  Dhara A. Patel, PhD, Yingjian You, BA, Guangming.

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Presentation on theme: "Interferon response and respiratory virus control are preserved in bronchial epithelial cells in asthma  Dhara A. Patel, PhD, Yingjian You, BA, Guangming."— Presentation transcript:

1 Interferon response and respiratory virus control are preserved in bronchial epithelial cells in asthma  Dhara A. Patel, PhD, Yingjian You, BA, Guangming Huang, BA, Derek E. Byers, MD, PhD, Hyun Jik Kim, MD, PhD, Eugene Agapov, PhD, Martin L. Moore, PhD, R. Stokes Peebles, MD, Mario Castro, MD, MPH, Kaharu Sumino, MD, MPH, Adrian Shifren, MD, Steven L. Brody, MD, Michael J. Holtzman, MD  Journal of Allergy and Clinical Immunology  Volume 134, Issue 6, Pages e7 (December 2014) DOI: /j.jaci Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Viral level and IFN response for IAV (A/WS/33 strain, MOI, 0.01) infection in hBEC cultures from asthmatic (n = 11) and nonasthmatic (n = 7) subjects. A, IAV RNA levels in cell lysate and supernatant at indicated times postinoculation (p.i.). B, IAV-induced cell toxicity marked by LDH release in cell supernatant. C, IFNB1, IFNL1, and IFNL2/3 mRNA levels. D, IFN-λ protein levels in cell supernatant. E, OAS1 and MX1 mRNA levels. #P < .05 for the set of values for IAV and UV-inactivated IAV (IAV-UV) in asthmatic and nonasthmatic subjects at each time point. The numbers in red indicate the fold-difference in the value for IAV in asthmatic versus nonasthmatic subjects at time points of statistical significance. LDH, Lactate dehydrogenase. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Viral level and IFN response for RSV (Long strain, MOI, 1) infection in hBEC cultures from asthmatic (n = 9) and nonasthmatic (n = 6) subjects. A, RSV RNA levels in cell lysate and supernatant. B, RSV-induced cell toxicity marked by LDH release in cell supernatant. C, IFNB1, IFNL1, and IFNL2/3 mRNA levels. D, IFN-λ protein levels in cell supernatant. E, OAS1 and MX1 mRNA levels. ###P < .001 and #P < .05 for the set of values for RSV and RSV-UV in asthmatic and nonasthmatic subjects at each time point. ***P < .001 and *P < .05 for the values for RSV in asthmatic versus nonasthmatic subjects over the time course of the experiment. The numbers in red indicate the fold-difference in the value for RSV in asthmatic versus nonasthmatic subjects at time points of statistical significance. LDH, lactate dehydrogenase; p.i., postinoculation. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Viral level and IFN response for RSV (A/2001/2-20 strain, MOI, 1) in hBEC cultures from asthmatic (n = 8) and nonasthmatic (n = 6) subjects. A, RSV RNA levels in cell lysate and supernatant. B, RSV-induced cell toxicity marked by LDH release in cell supernatant. C, IFNB1, IFNL1, and IFNL2/3 mRNA levels. D, IFN-λ protein levels in cell supernatant. E, OAS1 and MX1 mRNA levels. ###P < .001, ##P < .01, #P < .05 for the set of values for RSV and RSV-UV in asthmatic and nonasthmatic subjects at each time point. ***P < .001 and *P < .05 for the values for RSV in asthmatic versus nonasthmatic subjects over the time course of the experiment. The numbers in red indicate the fold-difference in the value for RSV in asthmatic versus nonasthmatic subjects at time points of statistical significance. LDH, lactate dehydrogenase; p.i., postinoculation. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 Correlations between levels of virus in cell supernatants versus levels of IFNB1, IFNL1, IFNL2/3, and OAS1 mRNA for hBECs obtained from all the asthmatic and nonasthmatic subjects and inoculated with IAV, RSV Long, or RSV A2001/2-20. Lines represent fits to a linear regression of log values. Values for viral levels represent mRNA copy number per microliter of cell supernatant. p.i., Postinoculation. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig E1 Titration of IAV inoculum in hTEC and hBEC cultures. Time course for levels of IAV PA RNA for MOI 0 to 1.0 in hTEC and hBEC cultures from nonasthmatic control subjects. p.i., Postinoculation. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig E2 Titration of IAV inoculum for optimal IFN expression in hTEC cultures. A, Time course for levels of IAV PA RNA for MOI 0 to 1.0. B, Corresponding levels of IFNB1 mRNA. C, Levels of IFNL1 mRNA. D, Levels of IFN-λ in cell supernatant. p.i., Postinoculation. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig E3 Viral level and IFN response for IAV (A/WS/33 strain, MOI, 0.01) infection in hBEC cultures from subjects with mild (n = 2), moderate (n = 6), and severe (n = 3) asthma as well as subjects without asthma (n = 7). A, IAV RNA levels in cell supernatant and lysate. B, IAV-induced cell toxicity marked by LDH release in cell supernatant. C, IFNB1, IFNL1, and IFNL2/3 mRNA levels. D, IFN-λ protein levels in cell supernatant. E, OAS1 and MX1 mRNA levels. Numbers in red represent fold-difference between asthmatic and nonasthmatic subjects at indicated time points. ***P < .001 and *P < .05 for comparisons over time for asthmatic versus nonasthmatic subjects. IAV-UV, UV-Inactivated IAV; LDH, lactate dehydrogenase; p.i., postinoculation. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig E4 Titration of RSV (Long strain) inoculum for optimal IFN expression in hTEC cultures. A, Time course for levels of RSV L RNA for MOI 0 to 1.0. B, Corresponding levels of IFNB1 mRNA. C, Levels of IFNL1 mRNA. D, Levels of IFN-λ protein in cell supernatant. p.i., Postinoculation. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Fig E5 Viral level and IFN response for RSV (Long strain, MOI, 1) infection in hBEC cultures from subjects with mild (n = 2), moderate (n = 5), and severe (n = 2) asthma as well as subjects without asthma (b = 6). A, RSV L RNA levels in cell supernatant and lysate. B, RSV-induced cell toxicity marked by LDH release in cell supernatant. C, IFNB1, IFNL1, and IFNL2/3 mRNA levels. D, IFN-λ protein levels in cell supernatant. E, OAS1 and MX1 mRNA levels. Numbers in blue represent fold-difference between moderate asthmatic and nonasthmatic subjects at indicated time points. ***P < .001 and *P < .05 for comparisons over time for asthmatic versus nonasthmatic subjects. LDH, Lactate dehydrogenase; p.i., postinoculation. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

11 Fig E6 Viral level and IFN response for RSV (A/2001/2-20 strain, MOI, 1) infection in hBEC cultures from subjects with mild (n = 2), moderate (n = 4), and severe (n = 2) asthma as well as subjects without asthma (n = 6). A, RSV RNA levels in cell supernatant and lysate. B, RSV-induced cell toxicity marked by LDH release in cell supernatant. C, IFNB1, IFNL1, and IFNL2/3 mRNA levels. D, IFN-λ protein levels in cell supernatant. E, OAS1 and MX1 mRNA levels. Numbers in red represent fold-difference between subjects with severe asthma and subjects without asthma at indicated time points. Numbers in orange represent fold-difference between subjects with mild asthma and subjects without asthma at indicated time points. **P < .01 and *P < .05 for comparisons over time for asthmatic versus nonasthmatic subjects. LDH, Lactate dehydrogenase; p.i., postinoculation. Journal of Allergy and Clinical Immunology  , e7DOI: ( /j.jaci ) Copyright © 2014 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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