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Quantitative evaluation of HIV and SIV co-receptor use with GHOST(3) cell assay.

Author

Editor

  • Tuofu Zhu

Summary, in English

An assay has been established for quantitative evaluation of lentivirus coreceptor use with the help of GHOST(3) cells. GHOST(3) cells were derived from the human osteosarcoma cell line, HOS, and have been engineered to stably express CD4 and one or another of the chemokine receptors CCR3, CCR5, CXCR4, CXCR6/STRL33/Bonzo, or the orphan receptor GPR15/BOB. The indicator cell line carries the HIV-2 long terminal repeat-driven green fluorescence protein (GFP) gene, which becomes activated upon infection with HIV or SIV. Viral entry is followed by Tat activation of transcription and GFP becomes expressed. Infected cells can be detected as early as 2 or 3 d after infection by simple fluorescence microscopic observation. The simplicity of the GHOST(3) cell system makes it particularly suitable for screening of a large number of isolates. In addition, the efficiency of co-receptor use can be accurately quantitated with flow cytometric analysis. Thus, the most efficiently used co-receptor of multitropic isolates can be determined. It is also possible to sensitively determine co-receptor switch of sequential isolates from the same individual.

Publishing year

2005

Language

English

Pages

333-342

Publication/Series

Human Retrovirus Protocols

Volume

2

Document type

Book chapter

Publisher

Humana Press

Topic

  • Microbiology in the medical area

Keywords

  • SIV
  • HIV
  • quantitative assay
  • co-receptor use
  • GFP
  • GHOST(3)

Status

Published

ISBN/ISSN/Other

  • ISSN: 1064-3745
  • ISSN: 1940-6029
  • ISBN: 978-1-58829-495-1