Department of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
Leidos Biomedical Research, Inc., Frederick, Maryland, USA.
J Clin Invest. 2020 Nov 2;130(11):5847-5857. doi: 10.1172/JCI138099.
BACKGROUNDHIV-1 viremia that is not suppressed by combination antiretroviral therapy (ART) is generally attributed to incomplete medication adherence and/or drug resistance. We evaluated individuals referred by clinicians for nonsuppressible viremia (plasma HIV-1 RNA above 40 copies/mL) despite reported adherence to ART and the absence of drug resistance to the current ART regimen.METHODSSamples were collected from at least 2 time points from 8 donors who had nonsuppressible viremia for more than 6 months. Single templates of HIV-1 RNA obtained from plasma and viral outgrowth of cultured cells and from proviral DNA were amplified by PCR and sequenced for evidence of clones of cells that produced infectious viruses. Clones were confirmed by host-proviral integration site analysis.RESULTSHIV-1 genomic RNA with identical sequences were identified in plasma samples from all 8 donors. The identical viral RNA sequences did not change over time and did not evolve resistance to the ART regimen. In 4 of the donors, viral RNA sequences obtained from plasma matched those sequences from viral outgrowth cultures, indicating that the viruses were replication competent. Integration sites for infectious proviruses from those 4 donors were mapped to the introns of the MATR3, ZNF268, ZNF721/ABCA11P, and ABCA11P genes. The sizes of the clones were estimated to be from 50 million to 350 million cells.CONCLUSIONThese findings show that clones of HIV-1-infected cells producing virus can cause failure of ART to suppress viremia. The mechanisms involved in clonal expansion and persistence need to be defined to effectively target viremia and the HIV-1 reservoir.FUNDINGNational Cancer Institute, NIH; Howard Hughes Medical Research Fellows Program, Howard Hughes Medical Institute; Bill and Melinda Gates Foundation; Office of AIDS Research; American Cancer Society; National Cancer Institute through a Leidos subcontract; National Institute for Allergy and Infectious Diseases, NIH, to the I4C Martin Delaney Collaboratory; University of Rochester Center for AIDS Research and University of Rochester HIV/AIDS Clinical Trials Unit.
尽管报告了药物依从性，并且当前的抗逆转录病毒治疗方案（ART）不存在耐药性，但组合抗逆转录病毒疗法（ART）仍无法抑制的 HIV-1 病毒血症通常归因于不完全的药物依从性和/或耐药性。我们评估了由于不可抑制的病毒血症（血浆 HIV-1 RNA 超过 40 拷贝/毫升）而被临床医生转介的个体，这些个体报告了对 ART 的依从性，并且当前的 ART 方案没有耐药性。
从至少 8 名病毒血症持续超过 6 个月的不可抑制病毒血症的供体中收集至少 2 个时间点的样本。通过 PCR 扩增从血浆和培养细胞的病毒生长物以及前病毒 DNA 中获得的 HIV-1 RNA 的单个模板，并进行测序以证明产生感染性病毒的细胞克隆的存在。通过宿主-前病毒整合位点分析来确认克隆。
从 8 名供体的血浆样本中均鉴定出具有相同序列的 HIV-1 基因组 RNA。相同的病毒 RNA 序列随时间不变，并且对 ART 方案没有产生耐药性。在 4 名供体中，从血浆中获得的病毒 RNA 序列与病毒生长培养物的序列相匹配，表明这些病毒具有复制能力。来自这 4 名供体的感染性前病毒的整合位点被映射到 MATR3、ZNF268、ZNF721/ABCA11P 和 ABCA11P 基因的内含子中。克隆的大小估计为 5000 万至 3.5 亿个细胞。
这些发现表明，产生病毒的 HIV-1 感染细胞的克隆可导致 ART 无法抑制病毒血症。需要确定涉及克隆扩增和持续存在的机制，以有效地靶向病毒血症和 HIV-1 储存库。
美国国立卫生研究院国家癌症研究所；霍华德·休斯医学研究所霍华德·休斯医学研究员计划；比尔和梅琳达·盖茨基金会；艾滋病研究办公室；美国癌症协会；美国国立卫生研究院通过 Leidos 分包合同资助罗切斯特大学艾滋病研究中心和罗切斯特大学艾滋病临床试验单位；国立过敏和传染病研究所，美国国立卫生研究院，I4C 马丁·德莱尼合作实验室。
