AIDSWEEKLY Plus, Monday, 12 May 1997
Daniel J. DeNoon, Senior Editor
A new assay speeds and simplifies tests for HIV infectivity and drug susceptibility.
The assay may even lead to a way to test patients' infectious viral burden, perhaps yielding previously unavailable prognostic data.
Currently, HIV infectivity is monitored by assessing viral p24 antigen production or by HeLa-CD4 plaque assays. Both tests are time consuming and yield limited data.
Now University of California, San Diego researchers Alain Gervaix and colleagues report the development of a new assay: a CEM cell line containing a plasmid encoding the green fluorescent protein (humanized S65T GFP) driven by the HIV-1 long terminal repeat (LTR) gene sequence. By monitoring increases in relative fluorescence by fluorescent microscopy, cytofluorimetry, and flow analysis, the cell line permits the rapid and direct evaluation of HIV infectivity and drug susceptibility.
"This technique enabled monitoring of HIV infection in real time, quantitation of infected cells over time, and determination of antiretroviral drug susceptibility easily and accurately," Gervaix et al. reported.
The researchers announced their findings in the Proceedings of the National Academy of Sciences ("A New Reporter Cell Line to Monitor HIV Infection and Drug Susceptibility In Vitro," PNAS, 1997;94:4653-8).
Gervaix et al. noted that the assay could be improved by providing the CEM-GFP cells with the CCR5 co-receptor that permits infection by macrophage-tropic, non-syncytium-inducing (NSI) HIV strains.
"The possibility of addressing infectious viral burden may have multiple practical implications for investigating viral replication, dynamics, and pathogenesis; for example, HIV induced apoptosis," the authors suggested. "Another potential application of the CEM-GFP reporter cell line could be to investigate cellular signaling pathways resulting in NF-kB activation."
This work was supported through the University of California, San Diego, Center for AIDS research development grant (National Institute of Allergy and Infectious Diseases 1 P30 AI 36214); Grants CA67394-02 from the National Institutes of Health and DK49618; Grants AI27670, AI38858, and AI29164 from the National Institutes of Health; grants from the Research Center for AIDS and HIV Infection of the San Diego Veterans Affairs Medical Center and by a training grant from the Swiss National Science Foundation.
The corresponding author for this study is Jaques Corbeil, Department of Medicine, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093-0679. Email: jcorbeil@ucsd.edu.
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