title: [The use of multi-color flow cytometry for identification of functional markers of nTregs in patients with severe asthma].
abstract: BACKGROUND
  At present, severe asthma is a particular clinical problem. An important role is attributed to dysfunction of nTreg subpopulations of lymphocytes in the pathogenesis of asthma. Therefore, the purpose of this study was to identify markers of nTreg cell function in patients with severe and mild to moderate asthma.
  
  
  METHODS
  The study included sixty patients with asthma (30 with severe and 30 with mild to moderate asthma). The control group comprised 30 healthy volunteers. The diagnosis of asthma was confirmed accordance with generally accepted recommendations (GINA 2008). nTreg immunophenotype CD4/CD25/CD127/FoxP3/GITR/CD152/CCR5/ /CCR7 was evaluated by multicolor flow cytometry.
  
  
  RESULTS
  We showed a significant reduction in the percentage of nTreg (76%) cells and the expression of CD152 (46.2%) in patients with severe asthma compared with mild-moderate asthma (85.5% and 86.7%; p 〈 0.05). It was observed that the transcription factor FoxP3 expression in nTreg cells positively correlated with FEV1 in patients with severe asthma (r = 0.53; p 〈 0.05). It was also found that the ratio nTregCCR5∗/TeffCCR5∗ was significantly reduced in patients with severe asthma (0.91) compared with mild-moderate (1.58) asthma and control groups (1.55; p 〈 0.001).
  
  
  CONCLUSIONS
  There are phenotypic differences in nTreg lymphocytes between patients with severe and mild-moderate asthma. This fact may confirm nTreg cell dysfunction and indicate that the potential markers (FoxP3, CD152, CCR5), can be used to monitor the effectiveness of treatment of bronchial asthma, especially severe disease.
authors.affiliation: Zakład Diagnostyki Laboratoryjnej, II Katedra Chorób Wewnętrznych, ul. Kopcińskiego 22, 90–153 Łódź. lukasz.kraszula@umed.lodz.pl
authors.forename: Lukasz
authors.initials: L
authors.lastname: Kraszula
authors.forename: Makandjou-Ola
authors.initials: M
authors.lastname: Eusebio
authors.forename: Maciej
authors.initials: M
authors.lastname: Kupczyk
authors.forename: Piotr
authors.initials: P
authors.lastname: Kuna
authors.forename: Mirosława
authors.initials: M
authors.lastname: Pietruczuk
mesh_terms: D000328:Adult; D000704:Analysis of Variance; D015703:Antigens, CD / Q000276:immunology*; D001249:Asthma / Q000276:immunology*; D015415:Biomarkers / Q000032:analysis; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D008297:Male; D008875:Middle Aged; D012129:Respiratory Function Tests; D012680:Sensitivity and Specificity; D012720:Severity of Illness Index; D018709:Statistics, Nonparametric; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*; D055815:Young Adult
chemical_list: D015703:Antigens, CD; D015415:Biomarkers