CG Dinucleotide Removal in Bioluminescent and Fluorescent Reporters Improves HIV-1 Replication and Reporter Gene Expression for Dual Imaging in Humanized Mice

Visualizing the transmission and dissemination of human immunodeficiency virus type 1 (HIV-1) in real time in humanized mouse models is a robust tool to investigate viral replication during treatments and in tissue reservoirs. However, the stability and expression of HIV-1 reporter genes are obstacl...

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Bibliographic Details
Published in:Journal of virology 2021-09, Vol.95 (19), p.e0044921-e0044921
Main Authors: Roy, Chandra N, Benitez Moreno, Mariana A, Kline, Chris, Ambrose, Zandrea
Format: Article
Language:English
Subjects:
Animals
CD4-Positive T-Lymphocytes - virology
Dinucleoside Phosphates - metabolism
Gene Expression
Genes, Reporter
HIV Infections - virology
HIV-1 - genetics
HIV-1 - physiology
Humans
Luciferases - genetics
Luminescent Measurements
Luminescent Proteins - genetics
Mice
Microbial Pathogenesis
Optical Imaging
Pathogenesis and Immunity
Viremia
Virus Replication
Whole Body Imaging
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Summary:Visualizing the transmission and dissemination of human immunodeficiency virus type 1 (HIV-1) in real time in humanized mouse models is a robust tool to investigate viral replication during treatments and in tissue reservoirs. However, the stability and expression of HIV-1 reporter genes are obstacles for long-term serial imaging . Two replication-competent CCR5-tropic HIV-1 reporter constructs were created that encode either nanoluciferase (nLuc) or a near infrared fluorescent protein (iRFP) upstream of . HIV-1 reporter virus replication and reporter gene expression was measured in cell culture and in humanized mice. While reporter gene expression correlated initially with plasma viremia, expression decreased after 4 to 5 weeks despite high plasma viremia. The reporter genes were codon optimized to remove cytosine/guanine (CG) dinucleotides, and new CO-nLuc and CO-iRFP viruses were reconstructed. Removal of CG dinucleotides in HIV-1 reporter viruses improved replication and reporter expression and . Both codon-optimized reporter viruses could be visualized during coinfection and reporter gene expression during treatment failure preceded detection of plasma viremia. While the dynamic range of CO-iRFP HIV-1 was lower than that of CO-nLuc HIV-1, both viruses could have utility in studying and visualizing HIV-1 infection in humanized mice. Animal models are important for studying HIV-1 pathogenesis and treatments. We developed two viruses each encoding a reporter gene that can be expressed in cells after infection. This study shows that HIV-1 infection can be visualized by noninvasive, whole-body imaging in mice with human immune cells over time by reporter expression. We improved reporter expression to reflect HIV-1 replication and showed that two viral variants can be tracked over time in the same animal and can predict failure of antiretroviral therapy to suppress virus.
ISSN:0022-538X
1098-5514
DOI:10.1128/JVI.00449-21