Laird Gregory M, Rosenbloom Daniel I S, Lai Jun, Siliciano Robert F, Siliciano Janet D
Department of Medicine, Johns Hopkins University School of Medicine, 733 N. Broadway, Baltimore, MD, 21205, USA.
Department of Biomedical Informatics, Columbia University Medical Center, New York, NY, 10032, USA.
Methods Mol Biol. 2016;1354:239-53. doi: 10.1007/978-1-4939-3046-3_16.
Combination antiretroviral therapy (cART) can reduce HIV-1 viremia to clinically undetectable levels. However, replication competent virus persists in a long-lived latent reservoir in resting, memory CD4(+) T cells. The latent reservoir in resting CD4(+) T cells is the major barrier to curing HIV-1 infection. The recent case of the Berlin patient has suggested that it may be possible to cure HIV-1 infection in certain situations. As efforts to cure HIV-1 infection progress, it will become critical to measure the latent reservoir in patients participating in clinical trials of eradication strategies. Our laboratory has developed a limiting dilution virus outgrowth assay that can be used to demonstrate the presence and persistence of latent HIV-1 in patients. Here we describe both the original and a simplified version of the quantitative virus outgrowth assay (QVOA) to measure the frequency of latently infected resting CD4(+) T cells with replication competent provirus in patients on suppressive cART.
联合抗逆转录病毒疗法（cART）可将HIV-1病毒血症降低至临床检测不到的水平。然而，具有复制能力的病毒持续存在于静息记忆CD4(+) T细胞的长寿潜伏库中。静息CD4(+) T细胞中的潜伏库是治愈HIV-1感染的主要障碍。最近的“柏林病人”案例表明，在某些情况下治愈HIV-1感染或许是可能的。随着治愈HIV-1感染的努力取得进展，对于参与根除策略临床试验的患者而言，检测潜伏库将变得至关重要。我们实验室已开发出一种极限稀释病毒增殖试验，可用于证明患者体内潜伏HIV-1的存在和持续情况。在此，我们描述了定量病毒增殖试验（QVOA）的原始版本和简化版本，以测量接受抑制性cART治疗的患者中携带具有复制能力前病毒的潜伏感染静息CD4(+) T细胞的频率。
