Enick P Nathan, Brooker Joseph P, Tumiotto Camille M, Staines Brittany T, Eron Joseph J, McMahon Deborah K, Gandhi Rajesh T, Mellors John W, Sobolewski Michele D
Division of Infectious Diseases, Department of Medicine, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
Division of Infectious Diseases, Department of Medicine, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
J Virus Erad. 2021 May 17;7(2):100043. doi: 10.1016/j.jve.2021.100043. eCollection 2021 Jun.
The quantitative viral outgrowth assay (qVOA) is the gold standard for measuring inducible, replication-competent HIV-1. Using MOLT4-R5 and SupT1-R5 cell lines instead of allogeneic blasts and HIV-1 RNA detection rather than p24 enzyme-immunoassay (EIA) has been proposed to improve the sensitivity of the qVOA. It is unclear, however, how these alternative approaches affect qVOA performance. We compared three qVOAs methods across 15 persons with HIV-1 on suppressive antiretroviral therapy and found that the MOLT4-R5 method yielded a significantly higher proportion of p24-positive wells (42%) than both the allogeneic blast (29%) and SupT1-R5 (32%) assays. Additionally, 5 of 7 qVOAs that were negative by p24 EIA showed viral outgrowth by HIV-1 RNA quantification (>10-fold increase within 7 days). These findings reveal the potential for underestimation of the latent, inducible reservoir by qVOA depending on the target cells used and the measure of viral outgrowth. Use of MOLT4-R5 cells with both p24 EIA and HIV-1 RNA to detect viral outgrowth was the most sensitive method.
定量病毒增殖测定（qVOA）是检测可诱导的、具有复制能力的HIV-1的金标准。有人提出，使用MOLT4-R5和SupT1-R5细胞系替代异基因胚细胞，并采用HIV-1 RNA检测而非p24酶免疫测定（EIA），以提高qVOA的灵敏度。然而，尚不清楚这些替代方法如何影响qVOA的性能。我们对15例接受抗逆转录病毒治疗且病毒得到抑制的HIV-1感染者的三种qVOA方法进行了比较，发现MOLT4-R5方法检测到的p24阳性孔比例（42%）显著高于异基因胚细胞检测法（29%）和SupT1-R5检测法（32%）。此外，7例p24 EIA检测为阴性的qVOA中有5例通过HIV-1 RNA定量检测显示有病毒增殖（7天内增加超过10倍）。这些发现揭示了根据所用靶细胞和病毒增殖检测方法的不同，qVOA可能低估潜伏的、可诱导的病毒储存库的情况。使用MOLT4-R5细胞并结合p24 EIA和HIV-1 RNA检测病毒增殖是最敏感的方法。
