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首页> 外文期刊>Journal of Clinical Microbiology >Identification and Characterization of HIV-1 Latent Viral Reservoirs In Peripheral Blood
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Identification and Characterization of HIV-1 Latent Viral Reservoirs In Peripheral Blood

机译：外周血HIV-1潜伏性病毒储库的鉴定与表征

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Plasma viral load and CD4 counts are effective for clinical monitoring, but they do not give a full representation of HIV-1 quasispecies in cellular reservoirs, the major repository of replication-competent HIV-1 in infected individuals. We sought to develop a diagnostic system that might stimulate the replication-competent HIV-1 reservoirs for enhanced clinical monitoring, including selection of antiretroviral regimens. Whole-blood samples from 45 HIV-infected individuals were collected into 1 ViraStim HIV-1 activation tube and 1 EDTA tube. Samples were tested for viral load and cell type-specific HIV-1 replication. Further, 7 matched activatedonactivated samples were sequenced using the Trugene HIV-1 genotyping kit. The percentage of patients with replication-competent virus in peripheral blood mononuclear cells (PBMCs) varied, depending on the baseline plasma viral load in the EDTA tubes. Six out of 24 patients with a starting plasma viral load of <20 copies/ml (cp/ml), 6 out of 8 patients with starting viral loads of >20 and <1,000 cp/ml, and 8 out of 13 patients with starting viral loads of >1,000 all showed increases in viral replication of >5-fold. These increases came from cellular reservoirs in blood as determined by simultaneous ultrasensitive subpopulation staining/hybridization in situ (SUSHI). When resistance genotypes in plasma from activation tubes were compared to those from EDTA tubes for 7 patients, all patients showed additional mutations in the activation tube, while 3 patients demonstrated additional genotypic resistance determinants. We show that HIV-1 viral replication can be stimulated directly from infected whole blood. The sequencing results showed that 3 of 7 cases demonstrated additional drug resistance following stimulation.
机译：血浆病毒载量和CD4计数可有效用于临床监测，但不能完全反映出细胞贮存器中HIV-1准种的完整情况，后者是感染个体中具有复制能力的HIV-1的主要贮存库。我们试图开发一种诊断系统，该系统可以刺激具有复制能力的HIV-1储库，以增强临床监测，包括选择抗逆转录病毒疗法。将来自45个HIV感染者的全血样品收集到1个ViraStim HIV-1激活管和1个EDTA管中。测试样品的病毒载量和特定细胞类型的HIV-1复制。此外，使用Trugene HIV-1基因分型试剂盒对7个匹配的激活/未激活样品进行了测序。外周血单个核细胞（PBMC）中具有复制能力的病毒患者的百分比有所不同，具体取决于EDTA管中的基线血浆病毒载量。 24例患者的起始血浆病毒载量小于20拷贝/毫升（cp / ml）中的6例，8例患者的起始病毒载量大于20和<1,000 cp / ml的6例，13例起始患者的8种> 1,000的病毒载量均显示> 5倍的病毒复制增加。这些增加来自血液中的细胞储库，这是通过同时进行超敏亚群染色/原位杂交（SUSHI）确定的。当比较7例激活管血浆中的抗性基因型与EDTA血浆中抗性基因型时，所有患者在激活管中均显示出额外的突变，而3例患者表现出其他基因型耐药性决定因素。我们表明，HIV-1病毒复制可以直接从感染的全血中刺激。测序结果表明，7例中的3例在刺激后表现出额外的耐药性。

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