title: Monocytes harbour replication-competent, non-latent HIV-1 in patients on highly active antiretroviral therapy.
abstract: OBJECTIVE
  To determine whether HIV-1 can be recovered from blood monocytes as well as resting, memory CD4 T lymphocytes of patients on highly active antiretroviral therapy (HAART) with undetectable plasma viraemia and whether infection is active or latent.
  
  
  METHODS
  Five patients with plasma HIV-1-RNA levels of less than 500 copies/ml for at least 3 months and less than 50 copies/ml at the time of sampling were initially selected, followed by an additional five patients with viral loads of less than 50 copies/ml for 3 months or more.
  
  
  METHODS
  Monocytes were isolated from blood by plastic adherence, then further purified by a second adherence step or CD3 depletion before co-culture with CD8-depleted donor peripheral blood mononuclear cells. Virus isolates were examined for mutations conferring resistance to reverse transcriptase or protease inhibitors and for genotype. The highly purified monocytes were also analysed for the presence of proviral and unintegrated viral DNA and multiply spliced (MS) viral mRNA by polymerase chain reaction.
  
  
  RESULTS
  Virus was recovered from monocytes of five patients. Sequencing of the recovered viruses did not reveal multiple drug resistance, and was consistent with a non-syncytium-inducing/CCR5 phenotype. Proviral DNA was detectable in monocytes from all subjects, and unintegrated HIV-1 DNA and MS RNA was found in four out of five populations examined.
  
  
  CONCLUSIONS
  Recovery of replication-competent virus from some HAART patients indicates that monocytes can also harbour HIV-1. Detection of circular, viral DNA and spliced RNA, albeit at very low levels, in these cells suggests that their infection is recent and transcriptionally active rather than latent.
authors.affiliation: AIDS Pathogenesis Research, Macfarlane Burnet Centre for Medical Research, Melbourne, Australia. sonza@burnet.edu.au
authors.forename: S
authors.initials: S
authors.lastname: Sonza
authors.forename: H P
authors.initials: HP
authors.lastname: Mutimer
authors.forename: R
authors.initials: R
authors.lastname: Oelrichs
authors.forename: D
authors.initials: D
authors.lastname: Jardine
authors.forename: K
authors.initials: K
authors.lastname: Harvey
authors.forename: A
authors.initials: A
authors.lastname: Dunne
authors.forename: D F
authors.initials: DF
authors.lastname: Purcell
authors.forename: C
authors.initials: C
authors.lastname: Birch
authors.forename: S M
authors.initials: SM
authors.lastname: Crowe
mesh_terms: D023241:Antiretroviral Therapy, Highly Active; D015496:CD4-Positive T-Lymphocytes / Q000821:virology; D015658:HIV Infections / Q000097:blood / Q000188:drug therapy / Q000821:virology*; D015497:HIV-1 / Q000235:genetics / Q000502:physiology; D006801:Humans; D009000:Monocytes / Q000821:virology*; D016662:Virus Integration; D017735:Virus Latency; D014779:Virus Replication