A computer-based, image-analysis method to quantify HIV-1 infection in a single-cycle infectious center assay.

TitleA computer-based, image-analysis method to quantify HIV-1 infection in a single-cycle infectious center assay.
Publication TypeJournal Article
Year of Publication2006
AuthorsDay JR, Martínez LE, Sásik R, Hitchin DL, Dueck ME, Richman DD, Guatelli JC
JournalJ Virol Methods
Volume137
Issue1
Pagination125-33
Date Published2006 Oct
ISSN0166-0934
KeywordsAnti-HIV Agents, Antibodies, Monoclonal, beta-Galactosidase, Genes, Reporter, HeLa Cells, HIV Antibodies, HIV Long Terminal Repeat, HIV-1, Humans, Image Processing, Computer-Assisted, Mutation, Neutralization Tests, Nevirapine, Receptors, CCR5, Receptors, CXCR4, Software
Abstract

Rapid and precise quantitation of the infectivity of HIV is important for molecular virologic studies as well as for measuring the activities of antiviral drugs and neutralizing antibodies. In the present study, an indicator cell line and image-analysis software were used to quantify HIV infectivity. Cells of the P4R5 line, which express the receptors for HIV infection as well as beta-galactosidase under the control of the HIV-1 long terminal repeat, were infected either with CXCR4- or CCR5-using viruses, including primary isolates, then stained 2 days later with X-gal to turn infected cells blue. Digital images of monolayers of the infected cells were captured using a high resolution CCD video camera and a macro video zoom lens. A software program was developed to process the images and to count the blue-stained foci of infection. The assay was applied to assess the infectivity of site-directed viral mutants, and to measure the activity of antiviral drugs and neutralizing antibody. The results indicate that the described method allows for the rapid quantitation of infected cells over a wide range of viral inocula with reproducibility, accuracy and relatively low cost.

DOI10.1016/j.jviromet.2006.06.019
Alternate JournalJ. Virol. Methods
PubMed ID16876264
Grant ListAI 36214 / AI / NIAID NIH HHS / United States
AI038201 / AI / NIAID NIH HHS / United States
AI043638 / AI / NIAID NIH HHS / United States
AI047745 / AI / NIAID NIH HHS / United States
AI27670 / AI / NIAID NIH HHS / United States
AI29164 / AI / NIAID NIH HHS / United States