title: CD4+CD25+CD127- assessment as a surrogate phenotype for FOXP3+ regulatory T cells in HIV-1 infected viremic and aviremic subjects.
abstract: BACKGROUND
  Although likely pivotal, the role of regulatory T cells (Tregs) in HIV pathogenesis remains elusive. This can be partly explained by analytical issues regarding their phenotypic identification in clinical studies. Instead of intracellular FOXP3 staining, CD4+CD25+CD127- phenotype has been proposed as an alternative to identify Tregs in clinical samples. However, its use remains controversial in viremic patients. Therefore, the objective of the present study was to assess the correlation between frequencies of CD4+CD25+CD127- and CD4+CD25+FOXP3+ lymphocytes in viremic and matched aviremic HIV-infected patients.
  
  
  METHODS
  Peripheral blood was collected from HIV-1 infected patients. Eleven viremic patients (Viral Load > 40 copies/mL) were matched (age, sex, CD4+ cell number) with 8 aviremic patients under highly active antiretroviral therapy (HAART). Fresh whole blood was immediately stained to analyze by flow cytometry the correlation between CD4+CD25+CD127- and the reference phenotype CD4+CD25+FOXP3+ lymphocytes in the same tube (four color staining CD4/CD25/CD127/FOXP3 for concomitant analysis of cell surface and intracellular markers).
  
  
  RESULTS
  In both groups, no significant differences were observed when comparing CD4+CD25+CD127- and CD4+CD25+FOXP3+ cell frequencies. In line, a strong correlation between CD4+CD25+CD127- and CD4+CD25+FOXP3+ lymphocyte percentages was observed in the whole patient population (r: 0.948, P < 0.001) or each group separately: aviremic (r: 0.968, P < 0.001), viremic (r: 0.9, P < 0.001). Finally, we found that most CD4+FOXP3+ cells were indeed CD25+CD127-, both in viremic and aviremic groups (88.5% and 90.9%, respectively).
  
  
  CONCLUSIONS
  We observed that CD4+CD25+CD127- phenotype is a good and easy-to-perform surrogate identification strategy for FOXP3+ regulatory T cells in both viremic and aviremic HIV-1-infected subjects. Thus, it represents a useful tool for monitoring Tregs in clinical research studies based on large cohorts of patients prospectively monitored, including HIV-infected subjects.
authors.affiliation: Hospices Civils de Lyon, Hôpital Edouard Herriot, Laboratoire d'Immunologie, Lyon, F-69003, France.
authors.forename: Julien
authors.initials: J
authors.lastname: Saison
authors.forename: Julie
authors.initials: J
authors.lastname: Demaret
authors.forename: Fabienne
authors.initials: F
authors.lastname: Venet
authors.forename: Christian
authors.initials: C
authors.lastname: Chidiac
authors.forename: Christophe
authors.initials: C
authors.lastname: Malcus
authors.forename: Francoise
authors.initials: F
authors.lastname: Poitevin-Later
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Tardy
authors.forename: Tristan
authors.initials: T
authors.lastname: Ferry
authors.forename: Guillaume
authors.initials: G
authors.lastname: Monneret
mesh_terms: D023241:Antiretroviral Therapy, Highly Active; D015704:CD4 Antigens / Q000032:analysis*; D005434:Flow Cytometry; D051858:Forkhead Transcription Factors / Q000032:analysis; D015658:HIV Infections / Q000097:blood / Q000175:diagnosis* / Q000276:immunology*; D015497:HIV-1; D006801:Humans; D053645:Interleukin-2 Receptor alpha Subunit / Q000032:analysis*; D053699:Interleukin-7 Receptor alpha Subunit / Q000032:analysis*; D010641:Phenotype; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000378:metabolism; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*; D014766:Viremia
chemical_list: D015704:CD4 Antigens; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D053645:Interleukin-2 Receptor alpha Subunit; D053699:Interleukin-7 Receptor alpha Subunit