Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Comparative Study
. 2019 Jan;160(1):107-115.
doi: 10.1177/0194599818803584. Epub 2018 Oct 16.

Pathologic Fibroblasts in Idiopathic Subglottic Stenosis Amplify Local Inflammatory Signals

Robert J Morrison  1   2 , Nicolas-George Katsantonis  1 , Kevin M Motz  3 , Alexander T Hillel  3 , C Gaelyn Garrett  1 , James L Netterville  1 , Christopher T Wootten  1 , Susan M Majka  4 , Timothy S Blackwell  4   5 , Wonder P Drake  6 , Alexander Gelbard  1
Affiliations
Comparative Study

Pathologic Fibroblasts in Idiopathic Subglottic Stenosis Amplify Local Inflammatory Signals

Robert J Morrison et al. Otolaryngol Head Neck Surg. 2019 Jan.

Abstract

Objective: To characterize the phenotype and function of fibroblasts derived from airway scar in idiopathic subglottic stenosis (iSGS) and to explore scar fibroblast response to interleukin 17A (IL-17A).

Study design: Basic science.

Setting: Laboratory.

Subjects and methods: Primary fibroblast cell lines from iSGS subjects, idiopathic pulmonary fibrosis subjects, and normal control airways were utilized for analysis. Protein, molecular, and flow cytometric techniques were applied in vitro to assess the phenotype and functional response of disease fibroblasts to IL-17A.

Results: Mechanistically, IL-17A drives iSGS scar fibroblast proliferation ( P < .01), synergizes with transforming growth factor ß1 to promote extracellular matrix production (collagen and fibronectin; P = .04), and directly stimulates scar fibroblasts to produce chemokines (chemokine ligand 2) and cytokines (IL-6 and granulocyte-macrophage colony-stimulating factor) critical to the recruitment and differentiation of myeloid cells ( P < .01). Glucocorticoids abrogated IL-17A-dependent iSGS scar fibroblast production of granulocyte-macrophage colony-stimulating factor ( P = .02).

Conclusion: IL-17A directly drives iSGS scar fibroblast proliferation, synergizes with transforming growth factor ß1 to promote extracellular matrix production, and amplifies local inflammatory signaling. Glucocorticoids appear to partially abrogate fibroblast-dependent inflammatory signaling. These results offer mechanistic support for future translational study of clinical reagents for manipulation of the IL-17A pathway in iSGS patients.

Keywords: IL-17; IL-17A; fibroblast; iSGS; idiopathic subglottis stenosis; laryngotracheal stenosis; tracheal stenosis.

PubMed Disclaimer

Conflict of interest statement

Disclosures

Competing interests: Alexander T. Hillel, Olympus, USA— consultant.

Sponsorships or competing interests that may be relevant to content are disclosed at the end of this article.

Figures

Figure 1.
Figure 1.
Airway scar in iSGS does not show evidence of EMT. (A) E-cadherin mRNA expression of iSGS and controls (mean ± SD). (B) E-cadherin immunohistochemistry of iSGS and controls. (C) No co-localization of epithelial and mesenchymal markers. Ctrl, control; EMT, epithelial-to-mesenchymal transition; iSGS, idiopathic subglottic stenosis; ns, not significant; CDH1, E-cadheirn.
Figure 2.
Figure 2.
iSGS patients do not show elevation in circulating fibrocytes. (A) Flow cytometry of PBMCs for circulating fibrocytes. (B) No significant difference in level of circulating fibrocytes between iSGS and control PBMCs. FSC, forward scatter; FITC, fluorescein isothiocyanate; IGG1, immunoglobulin G1; iSGS, idiopathic subglottic stenosis; PBMC, peripheral blood mononuclear cell; SSC, side scatter; Col-1, Type 1 collagen.
Figure 3.
Figure 3.
Fibroblasts derived from airway scar in iSGS proliferate in response to IL-17A. Values are presented as mean ± SD. *P < .05. (A) Fibroblasts from controls (left) or IPF (right) cultured with IL-17A. iSGS fibroblasts from 3 unique donors: (B) iSGS fibroblasts cultured with and without IL-17A; (C) iSGS fibroblasts cultured with IL-17A, IL-17rAb, nothing, or IL-17A and IL-17rAb. Ctrl, control; iSGS, idiopathic subglottic stenosis; IPF, intrapulmonary fibrosis; IL-17A, interleukin 17A; IL17rAb, interleukin 17A receptor antibody.
Figure 4.
Figure 4.
TGF-β synergizes with IL-17A to drive fibroblast collagen production in iSGS. (A) Flow cytometry levels of intracellular Col-1 and fibronectin iSGS of fibroblasts cultured with IL-17A. (B) qPCR levels of TGF-b mRNA in iSGS airway scar and controls. (C) Flow cytometry levels of Col-1 in iSGS fibroblasts from 3 unique donors, cultured with control media, IL-17A, TGF-β, or IL-17A and TGF- β. Values are presented as mean ± SD. *P < .05. Col-1, type 1 collagen; Ctrl, control; FN, fibronectin; IL-17A, interleukin 17A; iSGS, idiopathic subglottic stenosis; qPCR, quantitative polymerase chain reaction; TGF-β, transforming growth factor β.
Figure 5.
Figure 5.
IL-17A induces myeloid chemokines and cytokine production from iSGS fibroblasts. ELISA analysis of IL-1, GM-CSF, and CCL2 levels of iSGS fibroblasts cultured with IL-17A, GC, or IL-17A and GC. Values are presented as mean ± SD. *P < .05. CCL2, chemokine ligand 2; ELISA, enzyme-linked immunosorbent assay; GC, glucocorticoid; GM-CSF, granulocyte-macrophage colony-stimulating factor; IL, interleukin; iSGS, idiopathic subglottic stenosis.
See this image and copyright information in PMC

References

    1. Maldonado F, Loiselle A, Depew ZS, et al. Idiopathic subglottic stenosis: an evolving therapeutic algorithm. Laryngoscope. 2014;124:498–503. - PubMed
    1. Gelbard A, Donovan DT, Ongkasuwan J, et al. Disease homogeneity and treatment heterogeneity in idiopathic subglottic stenosis. Laryngoscope. 2016;126:1390–1396. - PMC - PubMed
    1. Nouraei SA, Sandhu GS. Outcome of a multimodality approach to the management of idiopathic subglottic stenosis. Laryngoscope. 2013;123:2474–2484. - PubMed
    1. Mark EJ, Meng F, Kradin RL, Mathisen DJ, Matsubara O. Idiopathic tracheal stenosis: a clinicopathologic study of 63 cases and comparison of the pathology with chondromalacia. Am J Surg Pathol. 2008;32:1138–1143. - PubMed
    1. Wang H, Wright CD, Wain JC, Ott HC, Mathisen DJ. Idiopathic subglottic stenosis: factors affecting outcome after single-stage repair. Ann Thorac Surg. 2015;100:1804–1811. - PubMed

Publication types

MeSH terms