Centre for Virus Research, The Westmead Institute for Medical Research, The University of Sydney, Sydney, NSW 2145, Australia.
Centre for Virus Research, The Westmead Institute for Medical Research, The University of Sydney, Sydney, NSW 2145, Australia.
Cell Rep. 2017 Oct 17;21(3):813-822. doi: 10.1016/j.celrep.2017.09.081.
Latent replication-competent HIV-1 persists in individuals on long-term antiretroviral therapy (ART). We developed the Full-Length Individual Proviral Sequencing (FLIPS) assay to determine the distribution of latent replication-competent HIV-1 within memory CD4 T cell subsets in six individuals on long-term ART. FLIPS is an efficient, high-throughput assay that amplifies and sequences near full-length (∼9 kb) HIV-1 proviral genomes and determines potential replication competency through genetic characterization. FLIPS provides a genome-scale perspective that addresses the limitations of other methods that also genetically characterize the latent reservoir. Using FLIPS, we identified 5% of proviruses as intact and potentially replication competent. Intact proviruses were unequally distributed between T cell subsets, with effector memory cells containing the largest proportion of genetically intact HIV-1 proviruses. We identified multiple identical intact proviruses, suggesting a role for cellular proliferation in the maintenance of the latent HIV-1 reservoir.
潜伏的复制型 HIV-1 存在于长期接受抗逆转录病毒治疗（ART）的个体中。我们开发了全长个体前病毒序列（FLIPS）检测方法，以确定六位长期接受 ART 的个体中记忆性 CD4 T 细胞亚群中潜伏的复制型 HIV-1 的分布。FLIPS 是一种高效、高通量的检测方法，可扩增和测序接近全长（约 9kb）的 HIV-1 前病毒基因组，并通过遗传特征确定潜在的复制能力。FLIPS 提供了一种基因组规模的视角，解决了其他遗传特征分析潜伏储库方法的局限性。使用 FLIPS，我们发现 5%的前病毒是完整的且具有潜在的复制能力。完整的前病毒在 T 细胞亚群之间分布不均，效应记忆细胞中含有最多比例的遗传完整 HIV-1 前病毒。我们还发现了多个相同的完整前病毒，这表明细胞增殖在维持潜伏 HIV-1 储库中发挥了作用。
