Innate immune responses of airway epithelium to house dust mite are mediated through β-glucan–dependent pathways Amy T. Nathan, MD, Elizabeth A. Peterson,

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Presentation transcript:

Innate immune responses of airway epithelium to house dust mite are mediated through β-glucan–dependent pathways Amy T. Nathan, MD, Elizabeth A. Peterson, BA, Jamila Chakir, PhD, Marsha Wills-Karp, PhD Journal of Allergy and Clinical Immunology Volume 123, Issue 3, Pages 612-618 (March 2009) DOI: 10.1016/j.jaci.2008.12.006 Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 CCL20 levels in HDM-treated epithelium. A, CCL20 protein levels determined by means of ELISA in supernatant from 16HBE14o- cells treated with varying doses of HDM. ∗P < .001. B, Time course of CCL20 secretion from 16HBE14o- cells treated with HDM (100 μg/mL). ∗P < .05 and ∗∗P < .001. C, CCL20 message from 16HBE14o- cells treated with HDM. P = not significant. D, CCL20 protein levels in HDM-exposed 16HBE14o- cells pretreated with cycloheximide (CHX) at 100 μg/mL. ∗P < .001. E, CCL20 protein levels determined by means of ELISA in supernatant from HDM-treated primary human bronchial epithelial cells. P < .001. ctrl, Control. Journal of Allergy and Clinical Immunology 2009 123, 612-618DOI: (10.1016/j.jaci.2008.12.006) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Effect of protease inhibition on CCL20 secretion. HDM was pretreated with a chemical protease inhibitor cocktail (PI) for 1 hour or heat treated for 20 minutes at 60°C. CCL20 protein levels were measured in supernatants from 16HBE14o- cells exposed to HDM, protease inhibitor–treated HDM, heat-inactivated HDM, or purified Der p 1. ∗P < .001. ctrl, Control. Journal of Allergy and Clinical Immunology 2009 123, 612-618DOI: (10.1016/j.jaci.2008.12.006) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Role of TLRs in mediating HDM effects on epithelium. A, CCL20 levels were measured in 16HBE14o- cells exposed to HDM, Pam3Cys, or LPS. ∗P < .001. B, 16HBE14o- cells transfected with a NF-κB TATA Luc reporter plasmid were stimulated with HDM or TNF-α, and NF-κB activation was assessed by using a luciferase activity. ∗P < .001. ctrl, Control. Journal of Allergy and Clinical Immunology 2009 123, 612-618DOI: (10.1016/j.jaci.2008.12.006) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Induction of CCL20 production by various environmental antigens. 16HBE14o- cells were exposed to HDM, ambient urban Baltimore particulate matter (AUB), LPS, OVA, ragweed, cockroach antigen (Ag), Der p 1, or chitin, and subsequent CCL20 secretion was measured by means of ELISA. ∗P < .001. ctrl, Control. Journal of Allergy and Clinical Immunology 2009 123, 612-618DOI: (10.1016/j.jaci.2008.12.006) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Role of β-glucan structures in HDM-induced CCL20 production. A, CCL20 secretion from 16HBE14o- cells treated with HDM, zymosan (Zym), laminarin (Lam), curdlan (Curd), and a combination of these β-glucans. ∗P < .001 vs control and ††P < .001 vs HDM. B, Dose-dependent inhibition of HDM-induced CCL20 secretion by laminarin. ∗P < .001 vs control and ††P < .001 vs HDM. C, Decreased CCL20 production by 16HBE14o- cells exposed to HDM pretreated with a β-glucanase. ∗∗P < .001 vs control, †P < .001 vs HDM, and §P < .01 vs maleic acid plus HDM. D, CCL20 secretion from HDM-exposed 16HBE14o- cells pretreated with the Syk inhibitor piceatannol (4 μmol/L final concentration). †P < .001 vs HDM. ctrl, Control. Journal of Allergy and Clinical Immunology 2009 123, 612-618DOI: (10.1016/j.jaci.2008.12.006) Copyright © 2009 American Academy of Allergy, Asthma & Immunology Terms and Conditions