DOI: 10.1073/pnas.94.9.4653. A new reporter cell line to monitor HIV infection and drug susceptibility<i>in vitro</i>

A new reporter cell line to monitor HIV infection and drug susceptibilityin vitro

10.1073/pnas.94.9.4653

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Determination of HIV infectivityin vitroand 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.

Bibliography

Gervaix, A., West, D., Leoni, L. M., Richman, D. D., Wong-Staal, F., & Corbeil, J. (1997). A new reporter cell line to monitor HIV infection and drug susceptibilityinâvitro. Proceedings of the National Academy of Sciences, 94(9), 4653â4658.

Authors 6

Alain Gervaix (first)
Daniel West (additional)
Lorenzo M. Leoni (additional)
Douglas D. Richman (additional)
Flossie Wong-Staal (additional)
Jacques Corbeil (additional)

References 33 Referenced 211

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Dates

Type	When
Created	23 years ago (July 26, 2002, 10:31 a.m.)
Deposited	2 years, 4 months ago (April 22, 2023, 8:11 a.m.)
Indexed	1 month, 2 weeks ago (July 3, 2025, 3:21 p.m.)
Issued	28 years, 3 months ago (April 29, 1997)
Published	28 years, 3 months ago (April 29, 1997)
Published Online	28 years, 3 months ago (April 29, 1997)
Published Print	28 years, 3 months ago (April 29, 1997)

Funders 0

None

BibTeX

@article{Gervaix_1997, title={A new reporter cell line to monitor HIV infection and drug susceptibilityin vitro}, volume={94}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.94.9.4653}, DOI={10.1073/pnas.94.9.4653}, number={9}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Gervaix, Alain and West, Daniel and Leoni, Lorenzo M. and Richman, Douglas D. and Wong-Staal, Flossie and Corbeil, Jacques}, year={1997}, month=apr, pages={4653–4658} }

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