Quantitative evaluation of HIV and SIV co-receptor use with GHOST(3) cell assay.
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.
Vödrös
Dalma
D
Division of Medical Microbiology, Department of Laboratory Medicine, Lund University, Lund, Sweden.
Fenyö
Eva Maria
EM
D000818:Animals; D045744:Cell Line, Tumor; D005434:Flow Cytometry / Q000379:methods; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006678:HIV / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D019707:Receptors, Chemokine / Q000096:biosynthesis / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D018000:Receptors, Peptide / Q000378:metabolism; D015302:Simian Immunodeficiency Virus / Q000302:isolation & purification / Q000378:metabolism*; D014162:Transfection
C103539:GPR15 protein, human; D019707:Receptors, Chemokine; D043562:Receptors, G-Protein-Coupled; D018000:Receptors, Peptide; D049452:Green Fluorescent Proteins