  HIV-1 infection currently cannot be cured because the virus persists as
integrated proviral DNA in long-lived cells despite years of suppressive
antiretroviral therapy (ART). To characterize establishment, turnover, and
evolution of viral DNA reservoirs we deep-sequenced the p17gag region of the
HIV-1 genome from samples obtained after 3-18 years of suppressive ART from 10
patients. For each of these patients, whole genome deep-sequencing data of
HIV-1 RNA populations before onset of ART were available from 6-12 longitudinal
plasma samples spanning 5-8 years of untreated infection. This enabled a
detailed analysis of the dynamics and origin of proviral DNA during ART. A
median of 14% (range 0-42%) of the p17gag DNA sequences were overtly defective
due to G-to-A hypermutation. The remaining sequences were remarkably similar to
previously observed RNA sequences and showed no evidence of evolution over many
years of suppressive ART. Most sequences from the DNA reservoirs were very
similar to viruses actively replicating in plasma (RNA sequences) shortly
before start of ART. The results do not support persistent HIV-1 replication as
a mechanism to maintain the HIV-1 reservoir during suppressive therapy. Rather,
the data indicate that viral DNA variants are turning over as long as patients
are untreated and that suppressive ART halts this turnover.
