title: Whole genome deep sequencing of HIV-1 reveals the impact of early minor variants upon immune recognition during acute infection.
abstract: Deep sequencing technologies have the potential to transform the study of highly variable viral pathogens by providing a rapid and cost-effective approach to sensitively characterize rapidly evolving viral quasispecies. Here, we report on a high-throughput whole HIV-1 genome deep sequencing platform that combines 454 pyrosequencing with novel assembly and variant detection algorithms. In one subject we combined these genetic data with detailed immunological analyses to comprehensively evaluate viral evolution and immune escape during the acute phase of HIV-1 infection. The majority of early, low frequency mutations represented viral adaptation to host CD8+ T cell responses, evidence of strong immune selection pressure occurring during the early decline from peak viremia. CD8+ T cell responses capable of recognizing these low frequency escape variants coincided with the selection and evolution of more effective secondary HLA-anchor escape mutations. Frequent, and in some cases rapid, reversion of transmitted mutations was also observed across the viral genome. When located within restricted CD8 epitopes these low frequency reverting mutations were sufficient to prime de novo responses to these epitopes, again illustrating the capacity of the immune response to recognize and respond to low frequency variants. More importantly, rapid viral escape from the most immunodominant CD8+ T cell responses coincided with plateauing of the initial viral load decline in this subject, suggestive of a potential link between maintenance of effective, dominant CD8 responses and the degree of early viremia reduction. We conclude that the early control of HIV-1 replication by immunodominant CD8+ T cell responses may be substantially influenced by rapid, low frequency viral adaptations not detected by conventional sequencing approaches, which warrants further investigation. These data support the critical need for vaccine-induced CD8+ T cell responses to target more highly constrained regions of the virus in order to ensure the maintenance of immunodominant CD8 responses and the sustained decline of early viremia.
authors.affiliation: Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Henn
authors.forename: Christian L
authors.initials: CL
authors.lastname: Boutwell
authors.forename: Patrick
authors.initials: P
authors.lastname: Charlebois
authors.forename: Niall J
authors.initials: NJ
authors.lastname: Lennon
authors.forename: Karen A
authors.initials: KA
authors.lastname: Power
authors.forename: Alexander R
authors.initials: AR
authors.lastname: Macalalad
authors.forename: Aaron M
authors.initials: AM
authors.lastname: Berlin
authors.forename: Christine M
authors.initials: CM
authors.lastname: Malboeuf
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: Ryan
authors.forename: Sante
authors.initials: S
authors.lastname: Gnerre
authors.forename: Michael C
authors.initials: MC
authors.lastname: Zody
authors.forename: Rachel L
authors.initials: RL
authors.lastname: Erlich
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Green
authors.forename: Andrew
authors.initials: A
authors.lastname: Berical
authors.forename: Yaoyu
authors.initials: Y
authors.lastname: Wang
authors.forename: Monica
authors.initials: M
authors.lastname: Casali
authors.forename: Hendrik
authors.initials: H
authors.lastname: Streeck
authors.forename: Allyson K
authors.initials: AK
authors.lastname: Bloom
authors.forename: Tim
authors.initials: T
authors.lastname: Dudek
authors.forename: Damien
authors.initials: D
authors.lastname: Tully
authors.forename: Ruchi
authors.initials: R
authors.lastname: Newman
authors.forename: Karen L
authors.initials: KL
authors.lastname: Axten
authors.forename: Adrianne D
authors.initials: AD
authors.lastname: Gladden
authors.forename: Laura
authors.initials: L
authors.lastname: Battis
authors.forename: Michael
authors.initials: M
authors.lastname: Kemper
authors.forename: Qiandong
authors.initials: Q
authors.lastname: Zeng
authors.forename: Terrance P
authors.initials: TP
authors.lastname: Shea
authors.forename: Sharvari
authors.initials: S
authors.lastname: Gujja
authors.forename: Carmen
authors.initials: C
authors.lastname: Zedlack
authors.forename: Olivier
authors.initials: O
authors.lastname: Gasser
authors.forename: Christian
authors.initials: C
authors.lastname: Brander
authors.forename: Christoph
authors.initials: C
authors.lastname: Hess
authors.forename: Huldrych F
authors.initials: HF
authors.lastname: Günthard
authors.forename: Zabrina L
authors.initials: ZL
authors.lastname: Brumme
authors.forename: Chanson J
authors.initials: CJ
authors.lastname: Brumme
authors.forename: Suzane
authors.initials: S
authors.lastname: Bazner
authors.forename: Jenna
authors.initials: J
authors.lastname: Rychert
authors.forename: Jake P
authors.initials: JP
authors.lastname: Tinsley
authors.forename: Ken H
authors.initials: KH
authors.lastname: Mayer
authors.forename: Eric
authors.initials: E
authors.lastname: Rosenberg
authors.forename: Florencia
authors.initials: F
authors.lastname: Pereyra
authors.forename: Joshua Z
authors.initials: JZ
authors.lastname: Levin
authors.forename: Sarah K
authors.initials: SK
authors.lastname: Young
authors.forename: Heiko
authors.initials: H
authors.lastname: Jessen
authors.forename: Marcus
authors.initials: M
authors.lastname: Altfeld
authors.forename: Bruce W
authors.initials: BW
authors.lastname: Birren
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Walker
authors.forename: Todd M
authors.initials: TM
authors.lastname: Allen
mesh_terms: D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D014644:Genetic Variation; D016679:Genome, Viral / Q000235:genetics*; D055106:Genome-Wide Association Study; D056914:Genomic Structural Variation; D015658:HIV Infections / Q000276:immunology / Q000517:prevention & control / Q000821:virology*; D015497:HIV-1 / Q000235:genetics* / Q000276:immunology / Q000472:pathogenicity; D006801:Humans; D057131:Immune Evasion / Q000235:genetics / Q000276:immunology*; D020411:Oligonucleotide Array Sequence Analysis; D012367:RNA, Viral / Q000032:analysis; D017423:Sequence Analysis, RNA; D014765:Viral Vaccines / Q000276:immunology
chemical_list: D012367:RNA, Viral; D014765:Viral Vaccines