Cigarette smoke combined with Toll-like receptor 3 signaling triggers exaggerated epithelial regulated upon activation, normal T-cell expressed and secreted/CCL5.

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Cigarette smoke combined with Toll-like receptor 3 signaling triggers exaggerated epithelial regulated upon activation, normal T-cell expressed and secreted/CCL5 expression in chronic rhinosinusitis Moshe Yamin, PhD, Eric H. Holbrook, MD, Stacey T. Gray, MD, Rachel Harold, BA, Nicolas Busaba, MD, Avinash Sridhar, Katia J. Powell, BS, Daniel L. Hamilos, MD Journal of Allergy and Clinical Immunology Volume 122, Issue 6, Pages 1145-1153.e3 (December 2008) DOI: 10.1016/j.jaci.2008.09.033 Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 1 Baseline gene expression in PNECs from HCs and subjects with CRS. Symbols (HC, open symbols; CRS, solid symbols) indicate level of expression relative to the housekeeping gene GAPDH. Subjects with CRS showed increased expression of TNF-α (4-fold increase; P = .02) and GRO-α (16-fold increase; P = .004) relative to HCs. Journal of Allergy and Clinical Immunology 2008 122, 1145-1153.e3DOI: (10.1016/j.jaci.2008.09.033) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 2 Representative epithelial immunostaining for TNF-α, GRO-α, and RANTES in HCs and subjects with CRS. A, Subject with CRS showing predominantly epithelial staining for TNF-α. B, Subject with CRS showing epithelial and glandular immunostaining for GRO-α. C, Subject with CRS showing epithelial immunostaining for RANTES. Inserts in A and C show the same tissues immunostained with the secondary antibody alone. Similar patterns of immunostaining were found in HCs. Magnification ×200. Journal of Allergy and Clinical Immunology 2008 122, 1145-1153.e3DOI: (10.1016/j.jaci.2008.09.033) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 3 Intensity of PNEC immunostaining in HCs versus subjects with CRS (N = 9 per group) for TNF-α (A), GRO-α (B), and RANTES (C). No statistically significant differences in immunostaining were found in HCs versus subjects with CRS. Journal of Allergy and Clinical Immunology 2008 122, 1145-1153.e3DOI: (10.1016/j.jaci.2008.09.033) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 4 Inflammatory gene induction in HCs and subjects with CRS by CSE, TNF-α, LTA, LPS, CSE+TNF-α, CSE+LTA, and CSE+LPS. The ordinate shows fold increase in gene expression relative to GAPDH housekeeping gene. No statistically significant differences were found between HCs and subjects with CRS except increased GRO-α, which patterned the increased baseline GRO-α in subjects with CRS. Journal of Allergy and Clinical Immunology 2008 122, 1145-1153.e3DOI: (10.1016/j.jaci.2008.09.033) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 5 Inflammatory gene induction in HCs and subjects with CRS by CSE, dsRNA, and CSE+dsRNA. The ordinate shows fold increase in gene expression relative to GAPDH housekeeping gene. Statistically significant differences between HCs and subjects with CRS were confined to greater induction of HBD2 and RANTES by CSE+dsRNA, as shown. Journal of Allergy and Clinical Immunology 2008 122, 1145-1153.e3DOI: (10.1016/j.jaci.2008.09.033) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Fig 6 Induction of IFN-β, IFN-λ1, IFN-λ2/3, TLR-3, and IL-17R by CSE, dsRNA, and CSE+dsRNA in HCs and subjects with CRS. The ordinate shows fold increase in gene expression relative to GAPDH housekeeping gene. CSE alone suppressed induction of IFN-β in subjects with CRS; however, CSE+dsRNA induced equivalent expression of IFN-β and the other gene studied. Journal of Allergy and Clinical Immunology 2008 122, 1145-1153.e3DOI: (10.1016/j.jaci.2008.09.033) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

Viability of primary nasal epithelial cells after 1-hour pulse exposure to CSE assessed at 6 hours and 24 hours by Trypan blue dye exclusion. Viability after exposure to 5% CSE was 81% at 24 hours in both normal controls and patients with CRS. Journal of Allergy and Clinical Immunology 2008 122, 1145-1153.e3DOI: (10.1016/j.jaci.2008.09.033) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions

RANTES protein levels and correlation with RANTES mRNA expression in new set of PNEC culture supernatants. A, RANTES protein in PNEC culture supernatant measured by ELISA in 3 healthy controls and 3 subjects with CRS. B, Dot-plot of RANTES protein in PNEC culture supernatant versus RANTES mRNA relative expression in the same PNEC cultures (N = 8). Con, Control, unstimulated condition. Journal of Allergy and Clinical Immunology 2008 122, 1145-1153.e3DOI: (10.1016/j.jaci.2008.09.033) Copyright © 2008 American Academy of Allergy, Asthma & Immunology Terms and Conditions