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Substance P represents a novel first-line defense mechanism in the nose  Olivia Larsson, PhD, Lotta Tengroth, MSc, Yuan Xu, PhD, Rolf Uddman, MD, PhD,

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Presentation on theme: "Substance P represents a novel first-line defense mechanism in the nose  Olivia Larsson, PhD, Lotta Tengroth, MSc, Yuan Xu, PhD, Rolf Uddman, MD, PhD,"— Presentation transcript:

1 Substance P represents a novel first-line defense mechanism in the nose 
Olivia Larsson, PhD, Lotta Tengroth, MSc, Yuan Xu, PhD, Rolf Uddman, MD, PhD, Susanna Kumlien Georén, PhD, Lars-Olaf Cardell, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 141, Issue 1, Pages e3 (January 2018) DOI: /j.jaci Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Journal of Allergy and Clinical Immunology 2018 141, 128-136
Journal of Allergy and Clinical Immunology  , e3DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 1 Expression of TLRs in human nasal mucosa. Representative images from 1 healthy subject demonstrating expression of TLR3 (A), TLR4 (B), TLR7 (C), TLR9 (D), and a negative control (E), as well as β-III tubulin, in human nasal mucosa are shown. Biopsy specimens from 5 healthy subjects were stained. Green, TLRs; red, β-III tubulin; and yellow, colocalized. Scale bar = 50 μm. Journal of Allergy and Clinical Immunology  , e3DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 2 Expression of TLR7 in human nasal mucosa. Representative images from 1 healthy subject demonstrating expression of TLR7 (green) and epithelial cell adhesion molecule (EpCAM; red; A), β-III tubulin (red; B), and SP (blue) colocalization (light blue; C). Biopsy specimens from 6 healthy subjects were stained. Scale bar = 50 μm. Journal of Allergy and Clinical Immunology  , e3DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 3 Expression of TLRs in human trigeminal ganglia. Representative images from 1 healthy subject demonstrating expression of TLR3 (A), TLR4 (B), TLR7 (C), and TLR9 (D) in human trigeminal ganglia sections are shown. Biopsy specimens from 6 healthy subjects were stained. Scale bar = 50 μm. Journal of Allergy and Clinical Immunology  , e3DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 4 A-C, SP production by TGNs (n = 5; Fig 4, A), MNECs (n = 4-5; Fig 4, B), and HNECs (n = 4; Fig 4, C). *P < .05, **P < .01, and ***P < .001, comparing unstimulated versus stimulated cells. Black line = mean. D and E, TLR7 (green) in TGNs (neurofilament/red), DAPI (blue), and colocalization (yellow, n = 10). Scale bar = 50 μm. F, SP production after R-837 (30 μg/mL). Values are means ± SEMs (n = 4-5). G, SP production 15 minutes after R-837 (5 μg/mL; n = 5). ***P < .001. Black line = mean. Journal of Allergy and Clinical Immunology  , e3DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 5 Epithelial expression of TLRs after stimulation with SP. A, Unstained (black histogram), isotype (gray histogram), and NK1R (red histogram; n = 5). B-F, HNEC expression of TLR1 (Fig 5, B), TLR3 (Fig 5, C), TLR4 (Fig 5, D), TLR7 (Fig 5, E), and TLR9 (Fig 5, F) after stimulation with SP or vehicle for 30 minutes (n = 5). *P < .05 and **P < .01, comparing unstimulated versus stimulated cells. Values are means ± SEMs. Journal of Allergy and Clinical Immunology  , e3DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig 6 Epithelial expression of TLR4 after stimulation with SP. Unstimulated HNECs (A and C) and SP-stimulated HNECs (B and D; 100 nmol/L SP for 30 minutes) are shown, as representative images from 1 healthy subject demonstrating expression of TLR4 (green) and nucleus (blue; n = 5). Scale bar = 50 μm. Journal of Allergy and Clinical Immunology  , e3DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig 7 Inhibition of epithelial TLR expression after stimulation with SP. HNEC expression of TLR1 (A), TLR3 (B), TLR4 (C), TLR7 (D), and TLR9 (E) 30 minutes after SP stimulation (100 nmol/L) with or without prior incubation (3 minutes) with aprepitant (10 nmol/L; n = 5) is shown. *P < .05 and **P < .01, comparing all cells versus SP-stimulated cells. All values are means ± SEMs. Journal of Allergy and Clinical Immunology  , e3DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Fig E1 Flow cytometry gating strategy to detect TLR expression on HNECs. A, Example of cultured HNECs gated on their forward/side-scatter properties and TLR9 expression: unstained (gray histogram), unstimulated (red histogram), and SP stimulated (open histogram). B, Experimental designed color panel, including spectral overlap. AF, Alexa Fluor; APC, allophycocyanin. Journal of Allergy and Clinical Immunology  , e3DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

11 Fig E2 Epithelial expression of TLRs after stimulation with SP. HNEC expression of TLR2 (A), TLR5 (B), TLR6 (C), and TLR8 (D) after stimulation with SP (10/50/100 nmol/L) or vehicle for 30 minutes is shown (n = 3-4). All values were means ± SEMs. Journal of Allergy and Clinical Immunology  , e3DOI: ( /j.jaci ) Copyright © 2017 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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