title: Generation of a dual RT Env SHIV that is infectious in rhesus macaques.
abstract: BACKGROUND
  The best current animal model for HIV infection and evaluation of antiviral compounds is the Simian-human immunodeficiency virus (SHIV)/macaque system. There are multiple recombinant SHIVs available, but these viruses have limitations in evaluating combination drug strategies for prevention. Drug combinations that target reverse transcriptase (RT, either nRTI or nnRTI) and envelope (entry or fusion inhibitors) have to be tested separately, which does not permit the assessment of additive, synergistic, or antagonistic effects of ARV combinations. We describe construction of a dual SHIV containing both HIV RT and a CCR5-specific HIV envelope gene in a simian immunodeficiency virus backbone.
  
  
  METHODS
  The RT Env SHIV molecular clone was constructed using RT SHIV and SHIV162p3 sequences as templates to generate RT Env SHIV. RT Env SHIV was expanded in vitro in CD8-depleted macaque peripheral blood mononuclear cells (PBMC). Recombinant virus was used to infect a rhesus macaque (4.3 x 10(4) tissue culture infectious dose [TCID(50)], intravenously [IV]). A second passage in a macaque by IV transfer of 10 ml of blood obtained from the first infection was also done. The in vivo adapted virus stock from these macaques was used to produce high titer stocks in vitro and used to rectally infect an additional macaque.
  
  
  RESULTS
  Peak viral load reached 6 x 10(5) vRNA copies/ml in plasma in both IV-exposed macaques and remained detectable in the one animal for 16 weeks after infection. A viral stock (1.68 x 10(4) TCID(50)) derived from the second macaque passage has been produced in CD8-depleted rhesus PBMC and was successfully used to demonstrate mucosal transmission. The resulting RT Env SHIV retained the sensitivity to HIV RT and entry inhibitors of its parental viruses.
  
  
  CONCLUSIONS
  The objective of this study was to develop and characterize a SHIV recombinant virus for evaluating the efficacy of ART and microbicide products that target both HIV RT and/or Env-mediated entry. RT Env SHIV can productively infect macaques by both the IV and mucosal route, making it a valuable tool for transmission studies.
authors.affiliation: Laboratory Branch, Division of HIV/AIDS Prevention, National Center for HIV, STD, TB Prevention, Centers for Disease Control and Prevention, Atlanta, GA, USA. ajo9@cdc.gov
authors.forename: James M
authors.initials: JM
authors.lastname: Smith
authors.forename: Allison
authors.initials: A
authors.lastname: Dauner
authors.forename: Bin
authors.initials: B
authors.lastname: Li
authors.forename: Priya
authors.initials: P
authors.lastname: Srinivasan
authors.forename: James
authors.initials: J
authors.lastname: Mitchell
authors.forename: Michael
authors.initials: M
authors.lastname: Hendry
authors.forename: Dennis
authors.initials: D
authors.lastname: Ellenberger
authors.forename: Sal
authors.initials: S
authors.lastname: Butera
authors.forename: Ron A
authors.initials: RA
authors.lastname: Otten
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D005260:Female; D008253:Macaca mulatta; D008297:Male; D012194:RNA-Directed DNA Polymerase / Q000235:genetics*; D016097:Simian Acquired Immunodeficiency Syndrome / Q000635:transmission; D015302:Simian Immunodeficiency Virus / Q000201:enzymology / Q000235:genetics*; D014759:Viral Envelope Proteins / Q000235:genetics*
chemical_list: D014759:Viral Envelope Proteins; D012194:RNA-Directed DNA Polymerase