Volume 10 Supplement 1

Canadian Society of Allergy and Clinical Immunology Annual Scientific Meeting 2013

Open Access

Th17/Treg ratio derived using DNA methylation analysis discriminates allergen-induced early from dual asthmatic responses

  • Amrit Singh1Email author,
  • Masatsugu Yamamoto1,
  • Jian Ruan1,
  • Jung Young Choi1,
  • Gail M Gauvreau2,
  • Paul M O'Byrne2,
  • Sven Olek3,
  • Ulrich Hoffmueller3,
  • Christopher Carlsten4,
  • J Mark FitzGerald4,
  • Louis-Philippe Boulet5 and
  • Scott J Tebbutt1
Allergy, Asthma & Clinical Immunology201410(Suppl 1):A46


Published: 3 March 2014


Atopic allergic asthmatic individuals experience acute bronchoconstriction (early response) upon allergen exposure. Several hours after the initial exposure, some individuals exhibit a chronic late phase (dual responders, DRs) whereas others do not (early responders, ERs). The purpose of this study is to determine changes in Th17 and regulatory T (Treg) cell numbers and their associated gene expression profiles in whole blood between allergen-induced ERs and DRs.


14 participants with mild, atopic asthma (8 ERs and 6 DRs) underwent a cat allergen inhalation challenge as part of the AllerGen Clinical Investigator Collaborative. Whole blood was collected immediately prior to challenge (pre) and 2 hours post-challenge. DNA methylation analysis was used to measure the frequency of Th17, Treg, B and T cells (Epiontis, Germany). Whole blood transcriptome profiling was performed using Affymetrix GeneChip® Human Gene 1.0 ST Arrays. Statistical analysis was performed using R.


Sum of the T cell and B cell frequencies obtained using the methylation assays strongly correlated (r = 0.95) with the lymphocyte frequency obtained using a hematolyzer. Allergen inhalation did not significantly (p>0.05) change Th17, Treg, B and T cell counts between ERs and DRs. However, the Th17/Treg ratio was significantly (p=0.03) different between ERs and DRs post challenge. 199 genes positively correlated with Th17 cells at an FDR of 5%. 463 genes positively correlated with Treg cells at an FDR of 5%. Th17 genes were inversely correlated with Treg genes.


Th17/Treg ratio derived using DNA methylation analysis discriminates allergen-induced early from dual asthmatic responses. The inverse correlation between Th17 genes and Treg genes may be indicative of the inflammatory or suppressive phenotypes of these cells.

Authors’ Affiliations

James Hogg Research Centre, St. Paul’s Hospital, University of British Columbia
Department of Medicine, McMaster University
Epiontis GmbH
Vancouver Coastal Health Research Institute, Vancouver General Hospital
Centre de Pneumologie de L’Hopital, Université Laval


© Singh et al; licensee BioMed Central Ltd. 2014

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.