Co-culture of human bronchial fibroblasts and CD4+ T cells increases Th17 cytokine signature

PLoS One. 2013 Dec 5;8(12):e81983. doi: 10.1371/journal.pone.0081983. eCollection 2013.

Abstract

Background: Airway inflammation is an important characteristic of asthma and has been associated with airway remodelling and bronchial hyperreactivity. The mucosal microenvironment composed of structural cells and highly specialised extracellular matrix is able to amplify and promote inflammation. This microenvironment leads to the development and maintenance of a specific adaptive response characterized by Th2 and Th17. Bronchial fibroblasts produce multiple mediators that may play a role in maintaining and amplifying this response in asthma.

Objective: To investigate the role of bronchial fibroblasts obtained from asthmatic subjects and healthy controls in regulating Th17 response by creating a local micro-environment that promotes this response in the airways.

Methods: Human bronchial fibroblasts and CD4(+)T cells were isolated from atopic asthmatics and non-atopic healthy controls. CD4(+)T were co-cultured with bronchial fibroblasts of asthmatic subjects and healthy controls. RORc gene expression was detected by qPCR. Phosphorylated STAT-3 and RORγt were evaluated by western blots. Th17 phenotype was measured by flow cytometry. IL-22, IL17, IL-6 TGF-β and IL1-β were assessed by qPCR and ELISA.

Results: Co-culture of CD4(+)T cells with bronchial fibroblasts significantly stimulated RORc expression and induced a significant increase in Th17 cells as characterized by the percentage of IL-17(+)/CCR6(+) staining in asthmatic conditions. IL-17 and IL-22 were increased in both normal and asthmatic conditions with a significantly higher amount in asthmatics compared to controls. IL-6, IL-1β, TGF-β and IL-23 were significantly elevated in fibroblasts from asthmatic subjects upon co-culture with CD4(+)T cells. IL-23 stimulates IL-6 and IL-1β expression by bronchial fibroblasts.

Conclusion: Interaction between bronchial fibroblasts and T cells seems to promote specifically Th17 cells profile in asthma. These results suggest that cellular interaction particularly between T cells and fibroblasts may play a pivotal role in the regulation of the inflammatory response in asthma.

MeSH terms

  • Adult
  • Asthma / genetics*
  • Asthma / immunology
  • Asthma / pathology*
  • Bronchi / immunology
  • Bronchi / metabolism
  • Bronchi / pathology*
  • CD4-Positive T-Lymphocytes / immunology
  • CD4-Positive T-Lymphocytes / pathology*
  • Case-Control Studies
  • Cellular Microenvironment
  • Coculture Techniques
  • Female
  • Fibroblasts / immunology
  • Fibroblasts / pathology*
  • Gene Expression Regulation / immunology
  • Humans
  • Interleukin-17 / genetics
  • Interleukin-17 / immunology*
  • Interleukin-1beta / genetics
  • Interleukin-1beta / immunology
  • Interleukin-22
  • Interleukin-6 / genetics
  • Interleukin-6 / immunology
  • Interleukins / genetics
  • Interleukins / immunology
  • Male
  • Middle Aged
  • Nuclear Receptor Subfamily 1, Group F, Member 3 / genetics
  • Nuclear Receptor Subfamily 1, Group F, Member 3 / immunology
  • Phosphorylation
  • Primary Cell Culture
  • Receptors, CCR6 / genetics
  • Receptors, CCR6 / immunology
  • STAT3 Transcription Factor / genetics
  • STAT3 Transcription Factor / immunology
  • Signal Transduction
  • Transforming Growth Factor beta / genetics
  • Transforming Growth Factor beta / immunology

Substances

  • CCR6 protein, human
  • Interleukin-17
  • Interleukin-1beta
  • Interleukin-6
  • Interleukins
  • Nuclear Receptor Subfamily 1, Group F, Member 3
  • RORC protein, human
  • Receptors, CCR6
  • STAT3 Transcription Factor
  • STAT3 protein, human
  • Transforming Growth Factor beta

Grants and funding

This study was supported by the Canadian Institutes of Health Research (CIHR) and by the Natural Sciences and Engineering Research Council of Canada (NSERC). R.F. was a recipient of a fellowship from the Respiratory Health Network of the Fonds de la Recherche en Santé du Québec. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.