In vitro selection of variants of human immunodeficiency virus type 1 resistant to 3'-azido-3'-deoxythymidine and 2',3'-dideoxyinosine
In vitro selection of variants of human immunodeficiency virus type 1 resistant to 3'-azido-3'-deoxythymidine and 2',3'-dideoxyinosine
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体外筛选抗 3-叠氮基-3-脱氧胸苷和 2,3-二脱氧肌苷的 1 型人类免疫缺陷病毒变种
作者：
Q. Gao;Zhengxian Gu;M. Parniak;Xuguang Li;M. Wainberg
人类免疫缺陷病毒1型（HIV-1）的耐药变异已被体外筛选分离。MT-4细胞被实验室毒株（HIV-IIIB）或临床分离株(no。在含有亚有效浓度药物3'-叠氮-3'-脱氧胸腺嘧啶（AZT）和2',3'-二脱氧肌苷（ddI）的培养基中维持。在新鲜MT-4细胞上进行病毒繁殖的过程中，通过逐渐增加培养基中的药物浓度，我们能够分离出HIV-IIIB的变体和临床分离物187，它们对药物的耐药性增加了100倍。在没有药物压力的情况下，变异繁殖2个多月后，耐药表型保持稳定。然而，对一种药物耐药的变异对另一种药物几乎没有交叉耐药，这表明对这两种化合物耐药的遗传基础不同。利用聚合酶链反应（PCR）对这些核苷耐药变异进行基因型分析，之前的引物对与AZT耐药突变相对应。观察到基因型的异质性，已知pol密码子70和215突变发生在大多数由HIV-IIIB或临床菌株187产生的azt耐药变体中。然而，密码子67和219的突变较少被检测到，并且在四种对ddI耐药的变异中均未观察到这些变化。对其中两个菌株的逆转录酶编码区进行了克隆和测序研究，并证实了所获得的PCR数据。除了先前描述的负责AZT抗性的突变位点外，hiv - iiib抗性变体显示在位置108 （Val----Ala）和135 （Ile----Thr）发生突变，而菌株187的抗性变体在位置50 （Ile----Val）和135 （Ile----Val）发生突变。
Drug-resistant variants of human immunodeficiency virus type 1 (HIV-1) have been isolated by in vitro selection. MT-4 cells were infected with either a laboratory strain (HIV-IIIB) or a clinical isolate (no. 187) of HIV-1 and maintained in medium containing subeffective concentrations of the drugs 3'-azido-3'-deoxythymidine (AZT) and 2',3'-dideoxyinosine (ddI). By gradually increasing the drug concentration in the culture medium during propagation of the virus on fresh MT-4 cells, we were able to isolate variants of HIV-IIIB and clinical isolate 187 which showed up to 100-fold increases in resistance to the drugs. The drug resistance phenotypes remained stable after propagation of the variants in the absence of drug pressure for over 2 months. However, variants resistant to one drug showed little or no cross-resistance to the other, suggesting that the genetic bases for resistance to the compounds differed. Genotypic analysis of these nucleoside-resistant variants by polymerase chain reaction (PCR) with primer pairs previously shown to correspond to mutations responsible for resistance to AZT was also carried out. A heterogeneity of genotypes was observed, with known mutations at pol codons 70 and 215 occurring in most of the AZT-resistant variants generated from either HIV-IIIB or clinical strain 187. However, mutations in codons 67 and 219 were less frequently detected, and none of these changes were observed in each of four variants resistant to ddI. Cloning and sequencing studies of the reverse transcriptase coding region of two of the isolates were also performed and confirmed the PCR data that had been obtained. In addition to previously described mutation sites responsible for resistance to AZT, an HIV-IIIB-resistant variant was shown to be mutated at positions 108 (Val----Ala) and 135 (Ile----Thr), while a resistant variant of strain 187 was mutated at positions 50 (Ile----Val) and 135 (Ile----Val).
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影响因子：
56.9
作者：
PRESTON, BD;POIESZ, BJ;LOEB, LA
通讯作者：
LOEB, LA
影响因子：
56.9
作者：
LARDER, BA;DARBY, G;RICHMAN, DD
通讯作者：
RICHMAN, DD
干扰素-α 和 3-叠氮基-3-脱氧胸苷在小鼠体内具有高度协同作用，可预防急性逆转录病毒暴露后的病毒血症。
DOI：
--
发表时间：
1990
期刊：
Journal of acquired immune deficiency syndromes
影响因子：
--
作者：
Ruprecht,RM;Chou,TC;Chipty,F;Sosa,MG;Mullaney,S;O'Brien,L;Rosas,D
通讯作者：
Rosas,D
通过 RNA.RNA 杂交检测对齐多夫定和双脱氧肌苷敏感性降低的人类免疫缺陷病毒 1 型临床分离株。
DOI：
10.1073/pnas.88.8.3092
发表时间：
1991
影响因子：
11.1
作者：
Japour,AJ;Chatis,PA;Eigenrauch,HA;Crumpacker,CS
通讯作者：
Crumpacker,CS