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

• DOI: 10.1038/srep43231
• Publication Date: 2017-02-24T11:25:30Z
• Authors: Axel Fun; Hoi Ping Mok; Mark R. Wills; Andrew M. Lever
• Keywords: Latent Virus; Virus latency

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