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A highly reproducible quantitative viral outgrowth assay for the measurement of the replication-competent latent HIV-1 reservoir

Published version
Peer-reviewed

Type

Article

Change log

Authors

Fun, A 
Mok, HP 
Wills, MR 
Lever, AM 

Abstract

Cure of Human Immunodeficiency Virus (HIV) infection remains elusive due to the persistence of HIV in a latent reservoir. Strategies to eradicate latent infection can only be evaluated with robust, sensitive and specific assays to quantitate reactivatable latent virus. We have taken the standard peripheral blood mononuclear cell (PBMC) based viral outgrowth methodology and from it created a logistically simpler and more highly reproducible assay to quantify replication-competent latent HIV in resting CD4+ T cells, both increasing accuracy and decreasing cost and labour. Purification of resting CD4+ T cells from whole PBMC is expedited and achieved in 3 hours, less than half the time of conventional protocols. Our indicator cell line, SupT1-CCR5 cells (a clonal cell line expressing CD4, CXCR4 and CCR5) provides a readily available standardised readout. Reproducibility compares favourably to other published assays but with reduced cost, labour and assay heterogeneity without compromising sensitivity.

Description

Keywords

retrovirus, viral reservoirs

Journal Title

Scientific Reports

Conference Name

Journal ISSN

2045-2322
2045-2322

Volume Title

7

Publisher

Nature Publishing Group
Sponsorship
Medical Research Council (MR/N02043X/1)
Academy of Medical Sciences (unknown)
British HIV Association (BHIVA) (unknown)
Imperial College London (P44848)
This work was submitted on behalf ofthe UK Biomedical Research Centres CHERUB cooperative and was supported by the National Institutes for Health Research (NIHR, Cambridge Biomedical Research Centres), Medical Research Council (MR/N02043X/1) and the Academy of Medical Sciences.