Rainwater-Lovett Kaitlin, Ziemniak Carrie, Watson Douglas, Luzuriaga Katherine, Siberry George, Petru Ann, Chen YaHui, Uprety Priyanka, McManus Margaret, Ho Ya-Chi, Lamers Susanna L, Persaud Deborah
Department of Pediatrics-Infectious Diseases, Johns Hopkins University School of Medicine, Baltimore, MD, United States of America.
Department of Pediatrics, University of Maryland, Baltimore, MD, United States of America.
PLoS One. 2017 Feb 8;12(2):e0170548. doi: 10.1371/journal.pone.0170548. eCollection 2017.
The latent reservoir is a major barrier to HIV eradication. Reservoir size is emerging as an important biomarker to assess the likelihood of HIV remission in the absence of antiretroviral therapy (ART) and may be reduced by earlier initiation of ART that restricts HIV spread into CD4+ T cells. Reservoir size is traditionally measured with a quantitative viral outgrowth assay (QVOA) that induces replication-competent HIV production through in vitro stimulation of resting CD4+ T cells. However, the recent identification of replication-intact, non-induced proviral genomes (NIPG) suggests the QVOA significantly underestimates (by 62-fold) latent reservoir size in chronically-infected adults. Whether formation and persistence of Intact, NIPG is thwarted by early ART initiation and long-term virologic suppression in perinatal infection is unclear. Here, we show that the latent reservoir in 11 early treated, long-term suppressed perinatally infected children and adolescents was not inducible by QVOA and dominated by defective, NIPG. Single genome analysis of 164 NIPG from 232 million cultured resting CD4+ T cells revealed no replication-intact, near-full length sequences. Forty-three (26%) NIPG contained APOBEC3G-mediated hypermutation, 115 (70%) NIPG contained large internal deletions, one NIPG contained nonsense mutations and indels, and 5 (3%) NIPG were assigned as "Not Evaluable" due to multiple failed sequencing attempts that precluded further classification. The lack of replication competent inducible provirus and intact NIPG in this cohort indicate early, long-term ART of perinatal infection leads to marked diminution of replication-competent HIV-1 reservoirs, creating a favorable state towards interventions aimed at virologic remission.
潜伏病毒库是根除艾滋病病毒的主要障碍。病毒库大小正成为一种重要的生物标志物，用于评估在不进行抗逆转录病毒疗法（ART）的情况下艾滋病病毒缓解的可能性，并且早期开始ART限制艾滋病病毒传播到CD4+T细胞中可能会使其减小。传统上，病毒库大小是通过定量病毒增殖测定（QVOA）来测量的，该测定通过体外刺激静止的CD4+T细胞诱导具有复制能力的艾滋病病毒产生。然而，最近对复制完整、未诱导的前病毒基因组（NIPG）的鉴定表明，QVOA显著低估了（62倍）慢性感染成年人中的潜伏病毒库大小。围产期感染中完整的NIPG的形成和持续是否会因早期ART启动和长期病毒学抑制而受阻尚不清楚。在这里，我们表明，11名早期接受治疗、长期病毒受到抑制的围产期感染儿童和青少年中的潜伏病毒库不能被QVOA诱导，且以有缺陷的NIPG为主。对来自2.32亿个培养的静止CD4+T细胞的164个NIPG进行单基因组分析，未发现复制完整、接近全长的序列。43个（26%）NIPG包含载脂蛋白B mRNA编辑酶催化多肽样3G（APOBEC3G）介导的超突变，115个（70%）NIPG包含大的内部缺失，1个NIPG包含无义突变和插入缺失，5个（3%）NIPG由于多次测序失败而无法进一步分类，被指定为“不可评估”。该队列中缺乏具有复制能力的可诱导前病毒和完整的NIPG表明，围产期感染的早期长期ART导致具有复制能力的艾滋病病毒-1病毒库显著减少，为旨在实现病毒学缓解的干预措施创造了有利条件。
