Emergence of Protease Inhibitor Resistance–Associated Mutations in Plasma HIV-1 Precedes That in Proviruses of Peripheral Blood Mononuclear Cells by More Than a Year

Abstract

HIV-1 genotype assay using plasma viruses has been widely applied for detection of resistant viruses in infected individuals, whereas there are only a few reports about proviral genotype in peripheral blood mononuclear cells (PBMCs). To determine which sample, plasma or PBMC, should be used for early detection of drug-resistant viruses during antiretroviral treatment, we analyzed 275 plasma-derived and 211 PBMC-derived HIV-1 protease sequences obtained from HIV-1-infected patients during protease inhibitor (PI) therapy. In 70 of 107 pairs (65.4%) of plasma and PBMC samples taken from the same blood draws, the numbers of PI resistance-associated mutations in the plasma-derived genotype were different from those in the PBMC-derived genotype. Plasma viruses had more PI resistance-associated mutations than PBMC proviruses (P = 0.0004). Analysis of serial samples showed that plasma-derived genotype assay could detect primary mutations about 425 days earlier than PBMC-derived genotype when the plasma viral load was less than 10(4 )copies/mL. Our data suggest that genetic turnover of PBMC proviruses is slower than that of plasma viruses and that time lag between emergence of mutations in plasma-derived and PBMC-derived genotypes correlates inversely with viral load. Plasma viruses should be the material of choice for early detection of drug resistance during antiretroviral treatment.

Feasibility of Detecting Human Immunodeficiency Virus Type 1 Drug Resistance in DNA Extracted from Whole Blood or Dried Blood Spots

“…Since all patients in this study initiated HAART very recently and about half of the 11 patients in whom drug resistance mutations were detected still had a viral load of less than 5,000 copies/ml at the time of sampling, they can be considered early treatment failures. Although the number of patients is limited, a correlation between low viral load and a reduced capacity for the detection of resistance mutations in DNA compared to that of resistance mutations in RNA was observed, further strengthening the hypothesis of a relationship between duration of treatment failure and the ability to detect drug resistance in DNA, as was suggested by Bi et al (6).…”

“…Some studies reported more mutations in the plasma RNA (6,10,21,24), and others reported more mutations in the cellular DNA (14,34). Most of these discordances may be attributed to differences in the selection of the samples used for the evaluation.…”

“…Results of published reports in this regard are very inconsistent (6,10,14,21,24,34). The present study aims to investigate both the possibility of DNA sequencing from whole blood cells or from DBS and the usefulness of DNA sequencing for determination of drug resistance in patients from a resource-poor setting with a high level of HIV-1 subtype diversity.…”

In Vivo Evidence for Instability of Episomal Human Immunodeficiency Virus Type 1 cDNA

“…Our observation that wild-type proviruses are maintained, despite the emergence of drug resistance mutations in plasma (1,17,34,37,42). In one study, emergence of the M184V mutation was compared in genomic viral RNA and viral DNA; however, there was no attempt to distinguish the emergence of the mutations in the different species of viral DNA (37).…”

Prevalence of HIV type 1 drug resistance mutations in treatment-naïve and experienced patients from resource-limited settings with universal access to antiretroviral therapy: a survey in two small Brazilian cities

“…Since we wished to study all patients (regardless of treatment history), including those with PVL below detection limit, we chose to perform resistance testing by targeting HIV-1 DNA from the PBMC proviral compartment, instead of the most commonly used plasma HIV-1 RNA. Although studies have found that the detection of resistance mutations in the HIV-1 RNA plasma compartment may precede that in PBMC (Kaye et al 1995, Bi et al 2003, it has been shown that HIV DNA recovered from the proviral compartment can reliably be used in drug resistance genotyping (Devereux et al 2000, Sarmati et al 2003, Chew et al 2005, Parisi et al 2007, Vicenti et al 2007). In addition, proviral sequences can be useful to detect archived mutations that can compromise future therapeutic options, such as those selected by previously used regimens (Sarmati et al 2003, SG Parisi et al unpublished observations), and have been found to have greater sensitivity after treatment interruption in treatment experienced patients (Venturi et al 2002).…”