A Novel Assay to Measure the Magnitude of the Inducible Viral Reservoir in HIV-infected Individuals

摘要: Abstract Background Quantifying latently infected cells is critical to evaluate the efficacy of therapeutic strategies aimed at reducing size long-lived viral reservoir, but low frequency these makes this very challenging. Methods We developed TILDA ( Tat / rev Induced Limiting Dilution Assay) measure with inducible multiply-spliced HIV RNA, as transcripts are usually absent in induced upon reactivation. requires less than a million cells, does not require RNA extraction and can be completed two days. Findings In suppressed individuals on ART, we found median CD4+ T estimated by 24cells/million, which was 48 times more measured quantitative outgrowth assay, 6–27 frequencies harbouring DNA PCR-based assays. measurements strongly correlated most The latent reservoir lower subjects who initiated ART during early compared late stage infection (p=0.011). untreated disease, carrying largely exceeded actively producing demonstrating that majority transcriptionally silent even absence ART. Interpretations Our results suggest reproducible sensitive approach productively clinical settings. demonstrate represents substantial fraction all prior initiation. Research context manuscript, describe development novel assay measures magnitude main barrier eradication. This termed for Tat/rev Assay, only 10ml blood, necessitate nucleic acids, highly reproducible, covers wide dynamic range sizes As such, may represent an alternative existing assays used reservoir.