Otero Miguel, Nunnari Giuseppe, Leto Daniela, Sullivan Julie, Wang Feng-Xiang, Frank Ian, Xu Yan, Patel Charvi, Dornadula Geethanjali, Kulkosky Joseph, Pomerantz Roger J
The Dorrance H. Hamilton Laboratories, Center for Human Virology and Biodefense, Division of Infectious Diseases and Environmental Medicine, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
AIDS Res Hum Retroviruses. 2003 Dec;19(12):1097-103. doi: 10.1089/088922203771881194.
Dendritic cells (DCs) are potent antigen-presenting cells, and their physiological localization in tissues that interact with the external environment is important as a first barrier against pathogens such as human immunodeficiency virus type I (HIV-1). Several models have been proposed to explain the possible role of DCs as a reservoir for HIV-1 in patients on virally suppressive highly active antiretroviral therapy (HAART). However, the low yield of cell isolates has made this evaluation a difficult task. The present study analyzes whether peripheral blood DCs from HIV-1-infected individuals on virally suppressive HAART, with plasma HIV-1 RNA levels of less than 50 copies/ml, carry either HIV-1 provirus and/or HIV-1 virions. Peripheral blood DCs were isolated from a cohort of 10 HIV-1-seropositive men taking suppressive HAART. In five patients, plasmacytoid and myeloid dendritic cells were isolated to attempt to identify their respective roles in HIV-1 residual disease. Viral out-growth assays were performed in vitro, as well as gag and R/U5 polymerase chain reaction (PCR) amplification of viral RNA and DNA, respectively, from DC and peripheral blood mononuclear cell (PBMC) extracts. Fluorescence activated cell-sorting (FACS) data revealed cellular yields from 85.90 to 95.18%, of relatively pure DCs isolated from patients' PBMCs. Although HIV-1 RNA gag and DNA RU/5 were detected in all PBMC samples isolated from the patients, proviral DNA and viral RNA forms were not detected in any of the DC isolates. In addition, no replication-competent virus was demonstrated in DC coculture assays, while virus was isolated from each patients' CD8+ T-lymphocyte-depleted PBMC cocultures. Furthermore, HIV-1 gag proviral DNA was not detected in either plasmacytoid or myeloid DC subfractions. The current study suggests that in HIV-1-infected individuals treated with suppressive HAART, peripheral blood DCs do not carry HIV-1 proviral DNA or viral particles attached to their surface. These populations of peripheral blood DCs are likely not a major HIV-1 reservoir in patients on HAART with clinically undetectable plasma viral RNA.
树突状细胞（DCs）是强大的抗原呈递细胞，其在与外部环境相互作用的组织中的生理定位作为抵御诸如I型人类免疫缺陷病毒（HIV-1）等病原体的第一道屏障至关重要。已经提出了几种模型来解释DCs在接受病毒抑制性高效抗逆转录病毒疗法（HAART）的患者中作为HIV-1储存库的可能作用。然而，细胞分离物的低产量使得这项评估成为一项艰巨的任务。本研究分析了接受病毒抑制性HAART且血浆HIV-1 RNA水平低于50拷贝/毫升的HIV-1感染个体的外周血DCs是否携带HIV-1前病毒和/或HIV-1病毒粒子。从一组10名接受抑制性HAART的HIV-1血清阳性男性中分离外周血DCs。在5名患者中，分离出浆细胞样和髓样树突状细胞，试图确定它们在HIV-1残留疾病中的各自作用。进行了体外病毒生长测定，以及分别从DC和外周血单核细胞（PBMC）提取物中对病毒RNA和DNA进行gag和R/U5聚合酶链反应（PCR）扩增。荧光激活细胞分选（FACS）数据显示，从患者PBMC中分离出的相对纯的DCs的细胞产量为85.90%至95.18%。尽管在从患者分离的所有PBMC样本中检测到了HIV-1 RNA gag和DNA RU/5，但在任何DC分离物中均未检测到前病毒DNA和病毒RNA形式。此外，在DC共培养试验中未证明有复制能力的病毒，而从每个患者的CD8 + T淋巴细胞耗竭的PBMC共培养物中分离出了病毒。此外，在浆细胞样或髓样DC亚组分中均未检测到HIV-1 gag前病毒DNA。当前研究表明，在接受抑制性HAART治疗的HIV-1感染个体中，外周血DCs不携带HIV-1前病毒DNA或附着于其表面的病毒颗粒。在血浆病毒RNA临床检测不到的HAART患者中，这些外周血DCs群体可能不是主要的HIV-1储存库。
