Holodniy M, Mole L, Margolis D, Moss J, Dong H, Boyer E, Urdea M, Kolberg J, Eastman S
AIDS Research Center, Department of Veterans Affairs Medical Center, Palo Alto, California, USA.
J Virol. 1995 Jun;69(6):3510-6. doi: 10.1128/JVI.69.6.3510-3516.1995.
Eleven human immunodeficiency virus (HIV)-infected subjects on long-term zidovudine (ZDV) therapy had didanosine (ddI) added to their antiretroviral regimen. HIV RNA in plasma was quantitated by branched-DNA signal amplification assay. Peripheral blood mononuclear cell (PBMC) HIV viral DNA was quantitated by PCR. The relative amounts of wild-type (WT) sequence, ddI resistance-associated codon changes (reverse transcriptase [RT] gene codon 65 K-->R [RT K65R], RT 174V, RT I135K/T/V, and RT M184I/V), and ZDV resistance-associated codon change (RT T215Y/F) from HIV RNA in plasma and RT T215Y/F from PBMC viral DNA were determined by differential hybridization of PCR products from 10 of 11 subjects. All subjects had evidence of RT T215Y/F mutation in both RNA in plasma and PBMC DNA at baseline. Subjects with a mixture of WT and RT T215Y/F HIV RNA in plasma at baseline demonstrated a decline in RNA levels in plasma after the addition of ddI. However, after 6 months of ZDV-ddI therapy, WT HIV RNA in plasma was undetectable in all subjects who had demonstrated a mixture at baseline. Subjects with only RT T215Y/F RNA present in plasma at baseline remained so and demonstrated no decline in RNA levels in plasma. In all subjects, no significant changes in PBMC DNA viral load and RT T215Y/F or WT levels were seen. HIV RNA in plasma demonstrated a significantly higher RT T215Y/F mutant/WT ratio than that of PBMC viral DNA, both at baseline and after ZDV-ddI combination therapy in all subjects. No subjects developed mutations associated with ddI resistance at codons 65, 74, 135, and 184 during this study. This study suggests that determination of relative amounts of RT T215Y/F and WT species from HIV RNA in plasma at baseline may be predictive of virologic response during ZDV-ddI combination therapy.
11名接受长期齐多夫定（ZDV）治疗的人类免疫缺陷病毒（HIV）感染者在其抗逆转录病毒治疗方案中添加了去羟肌苷（ddI）。通过分支DNA信号扩增测定法定量血浆中的HIV RNA。通过聚合酶链反应（PCR）定量外周血单个核细胞（PBMC）中的HIV病毒DNA。通过对11名受试者中10名的PCR产物进行差异杂交，确定血浆中HIV RNA的野生型（WT）序列、与ddI耐药相关的密码子变化（逆转录酶[RT]基因密码子65 K→R [RT K65R]、RT 174V、RT I135K/T/V和RT M184I/V）以及与ZDV耐药相关的密码子变化（RT T215Y/F）的相对量，以及PBMC病毒DNA中的RT T215Y/F。所有受试者在基线时血浆RNA和PBMC DNA中均有RT T215Y/F突变的证据。基线时血浆中存在WT和RT T215Y/F HIV RNA混合物的受试者在添加ddI后血浆RNA水平下降。然而，在ZDV-ddI治疗6个月后，所有在基线时表现出混合物的受试者血浆中的WT HIV RNA均无法检测到。基线时血浆中仅存在RT T215Y/F RNA的受试者仍然如此，且血浆RNA水平没有下降。在所有受试者中，PBMC DNA病毒载量以及RT T215Y/F或WT水平均无显著变化。在所有受试者中，无论是基线时还是ZDV-ddI联合治疗后，血浆中的HIV RNA显示出的RT T215Y/F突变体/WT比值均显著高于PBMC病毒DNA。在本研究期间，没有受试者在密码子65、74、135和184处发生与ddI耐药相关的突变。这项研究表明，在基线时测定血浆中HIV RNA的RT T215Y/F和WT种类的相对量可能预测ZDV-ddI联合治疗期间的病毒学反应。
