KNOCKING-OUT CONCENTRATIONS OF HIV-1-SPECIFIC INHIBITORS COMPLETELY SUPPRESS HIV-1 INFECTION AND PREVENT THE EMERGENCE OF DRUG-RESISTANT VIRUS
KNOCKING-OUT CONCENTRATIONS OF HIV-1-SPECIFIC INHIBITORS COMPLETELY SUPPRESS HIV-1 INFECTION AND PREVENT THE EMERGENCE OF DRUG-RESISTANT VIRUS
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DOI：
10.1006/viro.1993.1513
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发表时间：
1993-10-01
期刊：
影响因子：
3.7
通讯作者：
DECLERCQ, E
中科院分区：
文献类型：
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作者：
BALZARINI, J;KARLSSON, A;DECLERCQ, E
用浓度为0.1 μg/ml的HIV-1特异性抑制剂羟乙氧基甲基苯基硫代胸腺嘧啶（HEPT）、四氢咪唑苯并二氮杂卓酮（TIBO）、奈韦拉平、吡啶酮、双（杂芳基）哌嗪（BHAP）和叔丁基二甲基硅基螺氨基氧硫醚二氧化物（TSAO）处理HIV-1感染细胞，在3 - 4次传代培养中，耐药病毒迅速突破。在0.5 - 1 μg/ml的药物浓度下，耐药病毒的出现被延迟。在这些条件下产生的耐药HIV-1菌株的遗传和表型特征，并表现出不同的敏感性对不同类别的HIV-1特异性抑制剂，这取决于其逆转录酶中的氨基酸取代。在BHAP和吡啶酮抗性突变HIV-1株的逆转录酶中发现了新的氨基酸取代，这是迄今为止尚未报道的。在2.5至10 μg/ml浓度下，即在50%有效浓度（EC 50）的100至250倍浓度下，HEPT、TIBO、奈韦拉平、吡啶酮和BHAP在15次传代培养后可防止病毒突破。相反，3′-叠氮基-2 ′，3 ′-双脱氧胸苷（AZT），即使以1000倍高的浓度给药（即，1.3μg/ml）的EC 50不能阻止二次传代后的病毒突破。当以“敲除”浓度（2.5-10 μg/ml）使用时，HIV-1特异性抑制剂显然可以从HIV-1感染的细胞培养物中清除病毒，如通过（i）缺乏病毒致细胞病变性，（ii）缺乏病毒特异性包膜糖蛋白表达，（iii）缺乏病毒p24抗原产生和（iv）细胞中明显缺乏前病毒DNA所证明的。此外，加入HIV-1感染的CEM细胞（包括合胞体）的未感染的CEM细胞培养物可通过高浓度的HIV-1特异性抑制剂处理而免受破坏，但AZT和DDI（2′，3 ′-双脱氧肌苷）不能。
Treatment of HIV-1-infected cells with the HIV-1-specific inhibitors hydroxyethoxymethylphenylthiothymine (HEPT), tetrahydroimidazobenzodiazepinones (TIBO), nevirapine, pyridinone, bis(heteroaryl)piperazines (BHAP), andtert-butyl-dimethylsilylspiroaminooxathioledioxide (TSAO) at a concentration of 0.1 μg/ml resulted in a rapid breakthrough of resistant virus within three to four subcultivations. At drug concentrations of 0.5 to 1 μg/ml, emergence of resistant virus was delayed. The drug-resistant HIV-1 strains that originated under these conditions were genetically and phenotypically characterized and showed differential sensitivities against the different classes of HIV-1-specific inhibitors depending on the amino acid substitutions in their reverse transcriptase. Novel amino acid substitutions were found in the reverse transcriptase of BHAP- and pyridinone-resistant mutant HIV-1 strains that had not been reported so far. At 2.5 to 10 μg/ml, that is at a concentration 100- to 250-fold higher than the 50% effective concentration (EC50), HEPT, TIBO, nevirapine, pyridinone, and BHAP prevented virus breakthrough after 15 subcultivations. In contrast, 3′-azido-2′,3′-dideoxythymidine (AZT), even when administered at a 1000-fold higher concentration (i.e., 1.3 μg/ml) than its EC50failed to prevent virus breakthrough after the second subcultivation. HIV-1-infected cell cultures could apparently be cleared from virus by the HIV-1-specific inhibitors when used at the "knocking-out" concentrations (2.5-10 μg/ml), as evidenced by (i) the lack of viral cytopathicity, (ii) the lack of virus-specific envelope glycoprotein expression, (iii) the lack of viral p24 antigen production, and (iv) the apparent absence of proviral DNA in the cells. Moreover, uninfected CEM cell cultures to which HIV-1-infected CEM cells (including syncytia) had been added were protected from destruction by high-concentration treatment with the HIV-1-specific inhibitors, but not with AZT and DDI (2′,3′-dideoxyinosine).