4th Conference on Retroviruses and Opportunistic Infections


Washington, DC - January 22-26, 1997

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LTR-GFP: a new reporter system to monitor HIV infection and anti-HIV drug efficacy.

Conf Retroviruses Opportunistic Infect 1997 Jan 22-26; 4th:67 (abstract no. 16)

Gervaix A, West D, Wong-Staal F, Richman D, Corbeil J; University of California, San Diego, CA.

Introduction: Determination of antiretroviral drug activity by monitoring reduction of production of p24 antigen is expensive, time-consuming and does not allow accurate quantitation of the number of infected cells over time.

Aim: To develop a new simple, rapid and direct method for monitoring HIV infection and for testing anti-HIV drugs.

Method: We have produced a stable T-cell line (CEM) containing a plasmid encoding for a green fluorescent protein (humanized S65T GFP) driven by a HIV-1 LTR and selected clones that display low constitutive background of fluorescence but high level of GFP expression upon infection with HIV.

Results: HIV infection induced a 2-3x log increase in fluorescence of infected cells over 3 to 4 days which was easily monitored by fluorescence microscopy, by cytofluometry and by flow cytometry. Addition of anti-HIV drugs at different multiplicities of infection permitted the accurate determination of susceptibilities of both RT and protease inhibitors. This technique also permited quantitation of infectivity by assessment of number of cells infected in a first round of infection.

Conclusion: The LTR-GFP reporter system is a very simple, rapid and direct methodology for monitoring HIV infection in real time and will enable faster and easier antiretroviral drug susceptibilty testing.

Keywords: AEGIS, HIV Infections, Luminescent Proteins, HIV Long Terminal Repeat, Anti-HIV Agents, Reverse Transcriptase Inhibitors, HIV Protease Inhibitors, Organothiophosphorus Compounds, Microscopy, Fluorescence, Cell Line, Flow Cytometry, green fluorescent protein, methamidophos, genetics, AIDS

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