Quantification of airway fibrosis in asthma by flow cytometry

Research output: Contribution to journalArticle

Authors

  • Andrew Reichard
  • Nicholas Wanner
  • Eric Stuehr
  • Mario Alemagno
  • Kelly Weiss
  • Kimberly Queisser
  • Serpil Erzurum
  • Kewal Asosingh

External Institution(s)

  • Cleveland Clinic Foundation

Details

Original languageEnglish (US)
Pages (from-to)952-958
Number of pages7
JournalCytometry Part A
Volume93
Issue number9
StatusPublished - Sep 2018
Peer-reviewedYes

Abstract

Airway fibrosis is a prominent feature of asthma, contributing to the detrimental consequences of the disease. Fibrosis in the airway is the result of collagen deposition in the reticular lamina layer of the subepithelial tissue. Myofibroblasts are the leading cell type involved with this collagen deposition. Established methods of collagen deposition quantification present various issues, most importantly their inability to quantify current collagen biosynthesis occurring in airway myofibroblasts. Here, a novel method to quantify myofibroblast collagen expression in asthmatic lungs is described. Single cell suspensions of lungs harvested from C57BL/6 mice in a standard house dust mite model of asthma were employed to establish a flow cytometric method and compare collagen production in asthmatic and non-asthmatic lungs. Cells found to be CD45αSMA+, indicative of myofibroblasts, were gated, and median fluorescence intensity of the anti-collagen-I antibody labeling the cells was calculated. Lung myofibroblasts with no, medium, or high levels of collagen-I expression were distinguished. In asthmatic animals, collagen-I levels were increased in both medium and high expressers, and the number of myofibroblasts with high collagen-I content was elevated. Our findings determined that quantification of collagen-I deposition in myofibroblastic lung cells by flow cytometry is feasible in mouse models of asthma and indicative of increased collagen-I expression by asthmatic myofibroblasts.

    Research areas

  • airway, collagen-I, flow cytometry, myofibroblast

Citation formats

APA

Reichard, A., Wanner, N., Stuehr, E., Alemagno, M., Weiss, K., Queisser, K., ... Asosingh, K. (2018). Quantification of airway fibrosis in asthma by flow cytometry. Cytometry Part A, 93(9), 952-958. https://doi.org/10.1002/cyto.a.23373

Harvard

Reichard, A, Wanner, N, Stuehr, E, Alemagno, M, Weiss, K, Queisser, K, Erzurum, S & Asosingh, K 2018, 'Quantification of airway fibrosis in asthma by flow cytometry', Cytometry Part A, vol. 93, no. 9, pp. 952-958. https://doi.org/10.1002/cyto.a.23373