Nonnucleoside reverse transcriptase inhibitor-resistant HIV is stimulated by efavirenz during early stages of infection.
Nonnucleoside reverse transcriptase inhibitor-resistant HIV is stimulated by efavirenz during early stages of infection.
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在感染早期阶段，依非韦伦会刺激非核苷类逆转录酶抑制剂耐药的艾滋病毒。
DOI：
10.1128/jvi.05116-11
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发表时间：
2011
影响因子：
5.4
通讯作者：
Dykes,Carrie
中科院分区：
文献类型：
--
作者：
Wang,Jiong;Zhang,Gang;Bambara,RobertA;Li,Dongge;Liang,Hua;Wu,Hulin;Smith,HannahM;Lowe,NicholasR;Demeter,LisaM;Dykes,Carrie
非核苷类逆转录酶抑制剂（NNRTIs）是人类免疫缺陷病毒1型（HIV-1）感染联合治疗（CART）中常用的有效抗病毒药物。耐药性的发展是CART的主要限制。具有NNRTI耐药突变K101E+G190S的逆转录酶（RT）基因型对依非韦伦（EFV）具有高度耐药性，并且可以在患者含EFV方案失败时发生。我们之前已经证明，具有K101E+G190S突变的病毒在存在EFV时比不存在EFV时更有效地复制。在这项研究中，我们评估了药物依赖性刺激的潜在机制，使用单循环细胞培养试验，在感染或病毒产生步骤中添加EFV。我们确定EFV在早期感染时刺激K101E+G190S病毒，而不影响病毒复制的后期步骤，例如增加病毒颗粒中活性RT的数量。此外，我们发现另一种NNRTI，奈韦拉平（NVP），在感染的早期阶段刺激K101E+G190S病毒的复制，类似于EFV，但最新的NNRTI， etravirine (ETR)，没有。我们还发现EFV刺激K101E+Y188L和K101E+V106I病毒，但不刺激K101E+L100I、K101E+K103N、K101E+Y181C或K101E+G190A病毒，这表明刺激是突变特异性的。逆转录中间产物的Real-time PCR显示，虽然药物不刺激负链转移，但确实刺激负链强链DNA合成。我们的研究结果表明，刺激最有可能通过NNRTIs刺激tRNA启动或延伸的机制发生。
Nonnucleoside reverse transcriptase inhibitors (NNRTIs) are potent and commonly prescribed antiviral agents used in combination therapy (CART) of human immunodeficiency virus type 1 (HIV-1) infection. The development of drug resistance is a major limitation of CART. Reverse transcriptase (RT) genotypes with the NNRTI resistance mutations K101E+G190S are highly resistant to efavirenz (EFV) and can develop during failure of EFV-containing regimens in patients. We have previously shown that virus with K101E+G190S mutations can replicate more efficiently in the presence of EFV than in its absence. In this study, we evaluated the underlying mechanism for drug-dependent stimulation, using a single-cycle cell culture assay in which EFV was added either during the infection or the virus production step. We determined that EFV stimulates K101E+G190S virus during early infection and does not affect late steps of virus replication, such as increasing the amount of active RT incorporated into virions. Additionally, we showed that another NNRTI, nevirapine (NVP), stimulated K101E+G190S virus replication during the early steps of infection similar to EFV, but that the newest NNRTI, etravirine (ETR), did not. We also showed that EFV stimulates K101E+Y188L and K101E+V106I virus, but not K101E+L100I, K101E+K103N, K101E+Y181C, or K101E+G190A virus, suggesting that the stimulation is mutation specific. Real-time PCR of reverse transcription intermediates showed that although the drug did not stimulate minus-strand transfer, it did stimulate minus-strand strong-stop DNA synthesis. Our results indicate that stimulation most likely occurs through a mechanism whereby NNRTIs stimulate priming or elongation of the tRNA.