title: A Murine Viral Outgrowth Assay to Detect Residual HIV Type 1 in Patients With Undetectable Viral Loads.
abstract: BACKGROUND
  Sensitive assays are needed for detection of residual human immunodeficiency virus (HIV) in patients with undetectable plasma viral loads to determine whether eradication strategies are effective. The gold standard quantitative viral outgrowth assay (QVOA) underestimates the magnitude of the viral reservoir. We sought to determine whether xenograft of leukocytes from HIV type 1 (HIV)-infected patients with undetectable plasma viral loads into immunocompromised mice would result in viral amplification.
  
  
  METHODS
  Peripheral blood mononuclear cells or purified CD4(+) T cells from HIV or simian immunodeficiency virus (SIV)-infected subjects with undetectable plasma viral loads were adoptively transferred into NOD.Cg-Prkdc(scid)Il2rg(tm1Wjl)/SzJ (NSG) mice. The mice were monitored for viremia following depletion of human CD8(+) T cells to minimize antiviral activity. In some cases, humanized mice were also treated with activating anti-CD3 antibody.
  
  
  RESULTS
  With this murine viral outgrowth assay (MVOA), we successfully amplified replication-competent HIV or SIV from all subjects tested, including 5 HIV-positive patients receiving suppressive antiretroviral therapy (ART) and 6 elite controllers or suppressors who were maintaining undetectable viral loads without ART, including an elite suppressor from whom we were unable to recover virus by QVOA.
  
  
  CONCLUSIONS
  Our results suggest that the MVOA has the potential to serve as a powerful tool to identify residual HIV in patients with undetectable viral loads.
authors.affiliation: Department of Molecular and Comparative Pathobiology.
authors.forename: Kelly A
authors.initials: KA
authors.lastname: Metcalf Pate
authors.affiliation: Department of Medicine, Center for AIDS Research, Johns Hopkins University School of Medicine, Baltimore, Maryland.
authors.forename: Christopher W
authors.initials: CW
authors.lastname: Pohlmeyer
authors.affiliation: Department of Medicine, Center for AIDS Research, Johns Hopkins University School of Medicine, Baltimore, Maryland.
authors.forename: Victoria E
authors.initials: VE
authors.lastname: Walker-Sperling
authors.affiliation: Department of Molecular and Comparative Pathobiology.
authors.forename: Jeremy B
authors.initials: JB
authors.lastname: Foote
authors.affiliation: Department of Molecular and Comparative Pathobiology.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Najarro
authors.affiliation: Department of Molecular and Comparative Pathobiology University of Pennsylvania School of Veterinary Medicine, Philadelphia.
authors.forename: Catherine G
authors.initials: CG
authors.lastname: Cryer
authors.affiliation: Department of Medicine, Center for AIDS Research, Johns Hopkins University School of Medicine, Baltimore, Maryland Institute IrsiCaixa, Institut d'Investigació en Ciències de la Salut Germans Trias i Pujol, Universitat Autònoma de Barcelona, Badalona, Spain.
authors.forename: Maria
authors.initials: M
authors.lastname: Salgado
authors.affiliation: Department of Molecular and Comparative Pathobiology.
authors.forename: Lucio
authors.initials: L
authors.lastname: Gama
authors.affiliation: Department of Molecular and Comparative Pathobiology.
authors.forename: Elizabeth L
authors.initials: EL
authors.lastname: Engle
authors.affiliation: Department of Molecular and Comparative Pathobiology.
authors.forename: Erin N
authors.initials: EN
authors.lastname: Shirk
authors.affiliation: Department of Molecular and Comparative Pathobiology.
authors.forename: Suzanne E
authors.initials: SE
authors.lastname: Queen
authors.affiliation: Department of Medicine, Center for AIDS Research, Johns Hopkins University School of Medicine, Baltimore, Maryland.
authors.forename: Stanley
authors.initials: S
authors.lastname: Chioma
authors.affiliation: Department of Molecular and Comparative Pathobiology.
authors.forename: Meghan S
authors.initials: MS
authors.lastname: Vermillion
authors.affiliation: Department of Molecular and Comparative Pathobiology.
authors.forename: Brandon
authors.initials: B
authors.lastname: Bullock
authors.affiliation: Department of Molecular and Comparative Pathobiology.
authors.forename: Ming
authors.initials: M
authors.lastname: Li
authors.affiliation: Department of Molecular and Comparative Pathobiology Cummings School of Veterinary Medicine at Tufts University, North Grafton, Massachusetts.
authors.forename: Claire E
authors.initials: CE
authors.lastname: Lyons
authors.affiliation: Department of Molecular and Comparative Pathobiology.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Adams
authors.affiliation: Department of Molecular and Comparative Pathobiology.
authors.forename: M Christine
authors.initials: MC
authors.lastname: Zink
authors.affiliation: Department of Molecular and Comparative Pathobiology.
authors.forename: Janice E
authors.initials: JE
authors.lastname: Clements
authors.affiliation: Department of Molecular and Comparative Pathobiology.
authors.forename: Joseph L
authors.initials: JL
authors.lastname: Mankowski
authors.affiliation: Department of Medicine, Center for AIDS Research, Johns Hopkins University School of Medicine, Baltimore, Maryland.
authors.forename: Joel N
authors.initials: JN
authors.lastname: Blankson
mesh_terms: D000818:Animals; D023241:Antiretroviral Therapy, Highly Active; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000821:virology; D004195:Disease Models, Animal; D015658:HIV Infections / Q000175:diagnosis* / Q000188:drug therapy; D015497:HIV-1 / Q000254:growth & development / Q000302:isolation & purification*; D006801:Humans; D007376:Interleukin-2 / Q000097:blood; D007963:Leukocytes, Mononuclear / Q000821:virology; D008251:Macaca; D008297:Male; D051379:Mice; D015302:Simian Immunodeficiency Virus / Q000254:growth & development / Q000302:isolation & purification; D019562:Viral Load; D014766:Viremia / Q000662:veterinary
chemical_list: D007376:Interleukin-2
keywords: HIV; SIV; cure; humanized mouse; quantitative viral outgrowth assay (QVOA)