A new reporter cell line to monitor HIV infection and drug susceptibility in vitro.
Determination of HIV infectivity in vitro and its inhibition by antiretroviral drugs by monitoring reduction of production of p24 antigen is expensive and time consuming. Such assays also do not allow accurate quantitation of the number of infected cells over time. To develop a simple, rapid, and direct method for monitoring HIV infection, we generated a stable T-cell line (CEM) containing a plasmid encoding the green fluorescent protein (humanized S65T GFP) driven by the HIV-1 long terminal repeat. Clones were selected that displayed low constitutive background fluorescence, but a high level of GFP expression upon infection with HIV. HIV-1 infection induced a 100- to 1,000-fold increase in relative fluorescence of cells over 2 to 4 days as monitored by fluorescence microscopy, cytofluorimetry, and flow cytometry. Addition of inhibitors of reverse transcriptase, protease, and other targets at different multiplicities of infection permitted the accurate determination of drug susceptibility. This technique also permitted quantitation of infectivity of viral preparations by assessment of number of cells infected in the first round of infection. In conclusion, the CEM-GFP reporter cell line provides a simple, rapid, and direct method for monitoring HIV infectivity titers and antiretroviral drug susceptibility of syncytium-inducing strains.
Gervaix
A
A
Department of Medicine, University of California San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0679, USA.
West
D
D
Leoni
L M
LM
Richman
D D
DD
Wong-Staal
F
F
Corbeil
J
J
D019380:Anti-HIV Agents / Q000494:pharmacology*; D002460:Cell Line / Q000821:virology; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D015699:HIV Envelope Protein gp120 / Q000302:isolation & purification; D016325:HIV Long Terminal Repeat; D017320:HIV Protease Inhibitors / Q000494:pharmacology; D015497:HIV-1 / Q000254:growth & development*; D008164:Luminescent Proteins / Q000235:genetics; D008826:Microbial Sensitivity Tests / Q000379:methods*; D019829:Nevirapine; D011725:Pyridines / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis; D018894:Reverse Transcriptase Inhibitors / Q000494:pharmacology; D019258:Saquinavir / Q000494:pharmacology; D013601:T-Lymphocytes / Q000166:cytology / Q000821:virology*; D015215:Zidovudine / Q000494:pharmacology
D019380:Anti-HIV Agents; D015699:HIV Envelope Protein gp120; D017320:HIV Protease Inhibitors; D008164:Luminescent Proteins; D011725:Pyridines; D011994:Recombinant Proteins; D018894:Reverse Transcriptase Inhibitors; D049452:Green Fluorescent Proteins; D015215:Zidovudine; D019829:Nevirapine; D019258:Saquinavir