Günthard H F, Wong J K, Ignacio C C, Guatelli J C, Riggs N L, Havlir D V, Richman D D
Department of Pathology, School of Medicine, University of California San Diego, La Jolla 92093-0679, USA.
J Virol. 1998 Mar;72(3):2422-8. doi: 10.1128/JVI.72.3.2422-2428.1998.
Potent antiretroviral therapy can reduce human immunodeficiency virus (HIV) in plasma to levels below the limit of detection for up to 2 years, but the extent to which viral replication is suppressed is unknown. To search for ongoing viral replication in 10 patients on combination antiretroviral therapy for up to 1 year, the emergence of genotypic drug resistance across different compartments was studied and correlated with plasma viral RNA levels. In addition, lymph node (LN) mononuclear cells were assayed for the presence of multiply spliced RNA. Population sequencing of HIV-1 pol was done on plasma RNA, peripheral blood mononuclear cell (PBMC) RNA, PBMC DNA, LN RNA, LN DNA, and RNA from virus isolated from PBMCs or LNs. A special effort was made to obtain sequences from patients with undetectable plasma RNA, emphasizing the rapidly emerging lamivudine-associated M184V mutation. Furthermore, concordance of drug resistance mutations across compartments was investigated. No evidence for viral replication was found in patients with plasma HIV RNA levels of <20 copies/ml. In contrast, evolving genotypic drug resistance or the presence of multiply spliced RNA provided evidence for low-level replication in subjects with plasma HIV RNA levels between 20 and 400 copies/ml. All patients failing therapy showed multiple drug resistance mutations in different compartments, and multiply spliced RNA was present upon examination. Concordance of nucleotide sequences from different tissue compartments obtained concurrently from individual patients was high: 98% in the protease and 94% in the reverse transcriptase regions. These findings argue that HIV replication differs significantly between patients on potent antiretroviral therapy with low but detectable viral loads and those with undetectable viral loads.
强效抗逆转录病毒疗法可将血浆中的人类免疫缺陷病毒（HIV）水平降低至检测限以下长达2年，但病毒复制被抑制的程度尚不清楚。为了探寻10名接受联合抗逆转录病毒疗法长达1年的患者中是否存在持续的病毒复制，研究了不同区室间基因型耐药性的出现情况，并将其与血浆病毒RNA水平相关联。此外，还检测了淋巴结（LN）单核细胞中多重剪接RNA的存在情况。对HIV-1 pol进行群体测序，检测对象包括血浆RNA、外周血单核细胞（PBMC）RNA、PBMC DNA、LN RNA、LN DNA以及从PBMC或LN中分离出的病毒的RNA。特别努力地从血浆RNA检测不到的患者中获取序列，重点关注迅速出现的与拉米夫定相关的M184V突变。此外还研究了不同区室间耐药性突变的一致性。血浆HIV RNA水平<20拷贝/ml的患者未发现病毒复制的证据。相比之下，在血浆HIV RNA水平介于20至400拷贝/ml之间的受试者中，不断演变的基因型耐药性或多重剪接RNA的存在为低水平复制提供了证据。所有治疗失败的患者在不同区室均显示出多重耐药性突变，检查时存在多重剪接RNA。从个体患者同时获得的不同组织区室的核苷酸序列一致性很高：蛋白酶区为98%，逆转录酶区为94%。这些发现表明，在接受强效抗逆转录病毒疗法、病毒载量低但可检测到的患者与病毒载量检测不到的患者之间，HIV复制存在显著差异。
