title: Persistence of replication-competent HIV in both memory and naive CD4 T cell subsets in patients on prolonged and effective HAART.
abstract: OBJECTIVE
  To investigate the phenotypic features of infected lymphocytes in patients on prolonged and effective highly active antiretroviral therapy (HAART).
  
  
  METHODS
  We examined highly purified subsets of memory and naive CD4 T lymphocytes for the presence of replication-competent virus.
  
  
  METHODS
  In 11 highly selected HAART-treated patients, we isolated highly purified CD45RO CD45RA CD4 T cells using a magnetic bead-based procedure. In some patients, a subsequent cell separation according to CD62L expression was performed. We quantified total viral DNA in freshly isolated T-cell subsets. To verify whether the virus was replication-competent, HIV RNA was measured in supernatants following cell activation.
  
  
  RESULTS
  HIV DNA was detectable in the CD45RO and CD45RA CD4 T-cell subsets in 100% and 90% of the patients tested, respectively. In central memory CD45ROCD62L, effector memory CD45RO+CD62L-, truly naive CD45RACD62L, and CD45RA+CD62L- CD4 T cells, HIV DNA was found in 100%, 55%, 88%, and 50% of the patients tested respectively. HIV DNA was significantly higher in the CD45RO fraction than in the CD45RA subset and in the CD45ROCD62L fraction than in the three other CD45RA/ROCD62L+/- subsets. Detectable HIV RNA was found in the culture supernatants of CD45RO and CD45RA CD4 T-cell subsets in 80% and 66% of the patients tested respectively, and in CD45ROCD62L, CD45RO+CD62L-, CD45RACD62L, and CD45RA+CD62L- CD4 T cells in 100%, 100%, 100% and 50% of the patients tested respectively.
  
  
  CONCLUSIONS
  In patients on prolonged and effective HAART, the pool of infected CD4 T lymphocytes consists predominantly of memory cells but also contains naive cells.
authors.affiliation: INSERM E 0109, Faculté de Médecine Paris Sud, Hôpital du Kremlin Bicêtre, Le Kremlin Bicêtre, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Lambotte
authors.forename: Audrey
authors.initials: A
authors.lastname: Demoustier
authors.forename: Marie Ghislaine
authors.initials: MG
authors.lastname: de Goër
authors.forename: Christine
authors.initials: C
authors.lastname: Wallon
authors.forename: Jacques
authors.initials: J
authors.lastname: Gasnault
authors.forename: Cécile
authors.initials: C
authors.lastname: Goujard
authors.forename: Jean François
authors.initials: JF
authors.lastname: Delfraissy
authors.forename: Yassine
authors.initials: Y
authors.lastname: Taoufik
mesh_terms: D023241:Antiretroviral Therapy, Highly Active / Q000379:methods*; D015496:CD4-Positive T-Lymphocytes / Q000821:virology*; D002908:Chronic Disease; D015658:HIV Infections / Q000188:drug therapy* / Q000276:immunology / Q000821:virology; D006801:Humans; D007156:Immunologic Memory / Q000276:immunology; D012367:RNA, Viral / Q000032:analysis; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000821:virology; D019562:Viral Load; D014779:Virus Replication
chemical_list: D012367:RNA, Viral