Laboratory of Virology, EA2968, University of Bordeaux 2, Bordeaux, France.
J Antimicrob Chemother. 2010 Mar;65(3):434-7. doi: 10.1093/jac/dkp473. Epub 2010 Jan 5.
Our aim was to analyse the evolution of HIV-1 2-long terminal repeat (2-LTR) circular DNA in vitro and ex vivo in the presence of raltegravir.
Twenty-five patients starting a raltegravir-based regimen were included. Total HIV-1 DNA and 2-LTR DNA were quantified at baseline and in follow-up samples up to month 12. The effect of raltegravir on the formation of 2-LTR circles was evaluated in HeLa P4 cells. The effect of raltegravir was also investigated by sequence analysis of the 2-LTR circle junctions.
Among 21 patients with undetectable 2-LTR DNA at baseline, 7 had detectable 2-LTR DNA during the follow-up. Three of four patients with detectable 2-LTR DNA at baseline had undetectable 2-LTR DNA during the follow-up (P = 0.27). The mean 2-LTR level increased significantly (+0.07 log(10)/month, P = 0.02) in raltegravir-treated patients, and a 2-LTR increase was also observed in raltegravir-treated HeLa P4 cells, with a peak at 3 days post-infection. 2-LTR DNA showed a high prevalence of deletions ex vivo (64.5%) and in vitro (50%) in the presence of raltegravir, which was not statistically different from the prevalence in untreated patients or cells.
In antiretroviral-experienced patients receiving raltegravir, 2-LTR DNA increased while total HIV-1 DNA decreased over time. The frequent rearrangements found in 2-LTR sequences warrant further investigations to determine the dynamics of evolution of unintegrated HIV-1 DNA.
本研究旨在分析拉替拉韦存在时 HIV-1 2 长末端重复(2-LTR)环状 DNA 的体外和体内演变。
本研究共纳入 25 例开始使用拉替拉韦方案的患者。在基线和随访样本中,我们检测了总 HIV-1 DNA 和 2-LTR DNA,并在 12 个月时进行了随访。我们在 HeLa P4 细胞中评估了拉替拉韦对 2-LTR 环形成的影响,并通过对 2-LTR 环连接区的序列分析,研究了拉替拉韦的作用。
在 21 例基线时 2-LTR DNA 不可检测的患者中,7 例在随访期间出现了 2-LTR DNA 可检测。在基线时 2-LTR DNA 可检测的 4 例患者中,有 3 例在随访期间的 2-LTR DNA 不可检测(P = 0.27)。拉替拉韦治疗患者的 2-LTR 水平显著升高(+0.07 log(10)/月,P = 0.02),并且在拉替拉韦处理的 HeLa P4 细胞中也观察到了 2-LTR 增加,在感染后 3 天达到峰值。在拉替拉韦存在的情况下,2-LTR DNA 在外周血(64.5%)和体外(50%)均存在大量缺失,与未治疗患者或细胞中的缺失率无统计学差异。
在接受拉替拉韦治疗的抗逆转录病毒治疗经验丰富的患者中,2-LTR DNA 随时间推移而增加,而总 HIV-1 DNA 则减少。在 2-LTR 序列中发现的频繁重排需要进一步研究,以确定未整合 HIV-1 DNA 的进化动态。