title: Transcriptional Changes of Blood Eosinophils After Methacholine Inhalation Challenge in Asthmatics.
abstract: BACKGROUND
  Methacholine challenge is commonly used within the asthma diagnostic algorithm. Methacholine challenge has recently been shown to induce airway remodelling in asthma via bronchoconstriction, without additional airway inflammation. We evaluated the effect of methacholine-induced bronchoconstriction on the peripheral whole-blood transcriptome.
  
  
  METHODS
  Fourteen males with adult-onset, occupational asthma, 26-77 years of age, underwent methacholine inhalation challenges. The concentration of methacholine eliciting a ≥20% fall in FEV1 (PC20) was determined. Blood was collected immediately prior to and two hours after challenge. Complete blood counts and leukocyte differentials were obtained. Transcriptome analysis was performed using Affymetrix GeneChip(®) Human Gene 1.0 ST arrays. Data were analyzed using robust LIMMA and SAM. The cell-specific Significance Analysis of Microarrays (csSAM) algorithm was used to deconvolute the gene expression data according to cell type.
  
  
  RESULTS
  Microarray pathway analysis indicated that inflammatory processes were differentially affected. CsSAM identified 1,559 transcripts differentially expressed (all down-regulated) between pre- and post-methacholine in eosinophils at a false discovery cutoff of 10%. Notable changes included the GOLGA5 and METTL2B genes and the protein ubiquitination and CCR3 pathways.
  
  
  CONCLUSIONS
  We demonstrated significant changes in the peripheral blood eosinophil-specific transcriptome of asthmatics two hours after methacholine challenge. CCR3 and protein ubiquitination pathways are both significantly down-regulated.
authors.affiliation: James Hogg Research Centre, St. Paul's Hospital, University of British Columbia, Vancouver, British Columbia, Canada. ; Institute for HEART+LUNG Health, Vancouver, BC, Canada. ; Department of Medicine, Division of Respiratory Medicine, University of British Columbia, Vancouver, BC, Canada.
authors.forename: Scott J
authors.initials: SJ
authors.lastname: Tebbutt
authors.affiliation: James Hogg Research Centre, St. Paul's Hospital, University of British Columbia, Vancouver, British Columbia, Canada. ; Institute for HEART+LUNG Health, Vancouver, BC, Canada.
authors.forename: Jian-Qing
authors.initials: JQ
authors.lastname: He
authors.affiliation: James Hogg Research Centre, St. Paul's Hospital, University of British Columbia, Vancouver, British Columbia, Canada. ; Institute for HEART+LUNG Health, Vancouver, BC, Canada.
authors.forename: Amrit
authors.initials: A
authors.lastname: Singh
authors.affiliation: James Hogg Research Centre, St. Paul's Hospital, University of British Columbia, Vancouver, British Columbia, Canada. ; Institute for HEART+LUNG Health, Vancouver, BC, Canada.
authors.forename: Casey P
authors.initials: CP
authors.lastname: Shannon
authors.affiliation: James Hogg Research Centre, St. Paul's Hospital, University of British Columbia, Vancouver, British Columbia, Canada. ; Institute for HEART+LUNG Health, Vancouver, BC, Canada.
authors.forename: Jian
authors.initials: J
authors.lastname: Ruan
authors.affiliation: Institute for HEART+LUNG Health, Vancouver, BC, Canada. ; Department of Medicine, Division of Respiratory Medicine, University of British Columbia, Vancouver, BC, Canada. ; Vancouver Coastal Health Research Institute, Vancouver General Hospital, Vancouver, BC, Canada.
authors.forename: Chris
authors.initials: C
authors.lastname: Carlsten
keywords: CCR3; asthma; gene expression; microarray; peripheral blood; protein ubiquitination