Aoki S, Yarchoan R, Thomas R V, Pluda J M, Marczyk K, Broder S, Mitsuya H
Division of Cancer Treatment, National Cancer Institute, National Institute of Health, Bethesda, MD 20892.
AIDS Res Hum Retroviruses. 1990 Nov;6(11):1331-9. doi: 10.1089/aid.1990.6.1331.
A rapid and simple method for determining the proviral DNA content in peripheral blood mononuclear cells (PBM) from patients with human immunodeficiency virus type 1 (HIV-1) infection was established by using the polymerase chain reaction (PCR) technique. We found that the majority of HIV proviral DNA copies detectable in unfractionated PBM resided in T cells, while B cells/monocytes contained lesser amounts of HIV DNA (93.9 +/- 3.5% for T cells vs. 6.1 +/- 3.5% for B cells/monocytes: p less than 0.05). When we compared the amount of HIV proviral DNA in PBM from 13 patients with AIDS or AIDS-related complex (ARC) before and during antiretroviral therapy with 2',3'-dideoxyinosine (ddI) which was given as an escalating dose in a Phase I clinical study, a significant decrease was observed in 9 of 12 evaluable patients receiving the drug for 8 to 14 weeks (p less than 0.02). The decrease appeared more pronounced in patients receiving relatively high doses of the drug. These data suggest that the quantitation of HIV viral DNA in PBM by PCR is feasible and may theoretically contribute to an overall monitoring of patients receiving experimental therapy. However, larger studies will be required to determine the sensitivity and specificity of this assay and further longitudinal studies will be essential.
通过聚合酶链反应（PCR）技术建立了一种快速简便的方法，用于测定1型人类免疫缺陷病毒（HIV-1）感染患者外周血单核细胞（PBM）中的前病毒DNA含量。我们发现，在未分离的PBM中可检测到的大多数HIV前病毒DNA拷贝存在于T细胞中，而B细胞/单核细胞中的HIV DNA含量较少（T细胞为93.9±3.5%，B细胞/单核细胞为6.1±3.5%：p<0.05）。在一项I期临床研究中，我们将13例艾滋病或艾滋病相关综合征（ARC）患者在接受抗逆转录病毒药物2',3'-双脱氧肌苷（ddI）治疗前和治疗期间外周血单核细胞中的HIV前病毒DNA量进行了比较，该药物以递增剂量给药，在12例可评估患者中，9例接受该药物治疗8至14周后观察到显著下降（p<0.02）。在接受相对高剂量药物的患者中，下降更为明显。这些数据表明，通过PCR定量检测PBM中的HIV病毒DNA是可行的，理论上可能有助于对接受实验性治疗的患者进行全面监测。然而，需要进行更大规模的研究来确定该检测方法的敏感性和特异性，进一步的纵向研究也至关重要。
