title: Hematopoietic precursor cells isolated from patients on long-term suppressive HIV therapy did not contain HIV-1 DNA.
abstract: BACKGROUND
  We address the key emerging question of whether Lin(-)/CD34(+) hematopoietic precursor cells (HPCs) represent an important latent reservoir of human immunodeficiency virus type 1 (HIV-1) during long-term suppressive therapy.
  
  
  METHODS
  To estimate the frequency of HIV-1 infection in bone marrow, we sorted Lin(-)/CD34(+) HPCs and 3 other cell types (Lin(-)/CD34(-), Lin(-)/CD4(+), and Lin(+)/CD4(+)) from 8 patients who had undetectable viral loads for 3-12 years. Using a single-proviral sequencing method, we extracted, amplified, and sequenced multiple single HIV-1 DNA molecules from these cells and memory CD4(+) T cells from contemporaneous peripheral blood samples.
  
  
  RESULTS
  We analyzed 100,000-870,000 bone marrow Lin(-)/CD34(+) HPCs from the 8 patients and found no HIV-1 DNA. We did isolate HIV-1 DNA from their bone marrow Lin(+)/CD4(+) cells that was genetically similar to HIV-1 DNA from lymphoid cells located in the peripheral blood, indicating an exchange of infected cells between these compartments.
  
  
  CONCLUSIONS
  The absence of infected HPCs provides strong evidence that the HIV-1 infection frequency of Lin(-)/CD34(+) HPCs from bone marrow, if it occurred, was <.003% (highest upper 95% confidence interval) in all 8 patients. These results strongly suggest that Lin(-)/CD34(+) HPCs in bone marrow are not a source of persistent HIV-1 in patients on long-term suppressive therapy.
authors.affiliation: Department of Diagnostics and Vaccinology, Swedish Institute for Communicable Disease Control, Karolinska Institutet, Solna, Sweden.
authors.forename: Lina
authors.initials: L
authors.lastname: Josefsson
authors.forename: Susanne
authors.initials: S
authors.lastname: Eriksson
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Sinclair
authors.forename: Terence
authors.initials: T
authors.lastname: Ho
authors.forename: Maudi
authors.initials: M
authors.lastname: Killian
authors.forename: Lorrie
authors.initials: L
authors.lastname: Epling
authors.forename: Wei
authors.initials: W
authors.lastname: Shao
authors.forename: Bradley
authors.initials: B
authors.lastname: Lewis
authors.forename: Peter
authors.initials: P
authors.lastname: Bacchetti
authors.forename: Lisa
authors.initials: L
authors.lastname: Loeb
authors.forename: Jeff
authors.initials: J
authors.lastname: Custer
authors.forename: Lauren
authors.initials: L
authors.lastname: Poole
authors.forename: Frederick M
authors.initials: FM
authors.lastname: Hecht
authors.forename: Sarah
authors.initials: S
authors.lastname: Palmer
mesh_terms: D018952:Antigens, CD34 / Q000235:genetics / Q000276:immunology; D001853:Bone Marrow / Q000276:immunology / Q000821:virology; D015496:CD4-Positive T-Lymphocytes / Q000821:virology; D015331:Cohort Studies; D004279:DNA, Viral / Q000235:genetics* / Q000302:isolation & purification*; D015658:HIV Infections / Q000276:immunology / Q000821:virology*; D015497:HIV-1 / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D006412:Hematopoietic Stem Cells / Q000276:immunology / Q000821:virology*; D006801:Humans; D010802:Phylogeny; D019562:Viral Load / Q000235:genetics
chemical_list: D018952:Antigens, CD34; D004279:DNA, Viral