MECHANISMS OF RESISTANCE IN CLINICAL ISOLATES OF HIV

HIV 临床分离株的耐药机制

• 批准号: 3143924
• 负责人: DOUGLAS D RICHMAN
• 金额: $ 27.33万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-09-30 至 1994-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3143924
• 关键词: 2'3' dideoxycytidine; 2'3' dideoxynucleoside; AIDS; AIDS therapy; CD4 molecule; HeLa cells; RNA directed DNA polymerase; antiAIDS agent; drug resistance; genetic manipulation; human immunodeficiency virus; laboratory mouse; nucleic acid probes; nucleic acid sequence; phospholipids; polymerase chain reaction; severe combined immunodeficiency; site directed mutagenesis; zidovudine

During the past year we have obtained over 60 isolates of HIV-1 from over 40 patients, many of whom have been on prolonged therapy with AZT (3'- azido, 3'-deoxythymidine, zidovudine) for AIDS or advanced AIDS-related complex. We have also developed a plaque reduction assay in a CD4-HeLa cell line that permits reproducible quantitative assays of drug sensitivity of these isolates. Fifty percent inhibitory (ID50) values of 18 isolates from untreated individuals ranged between 0.01muM and 0.05muM. In contrast, most isolates from patients who had received AZT for 6 months or more exhibited decreased sensitivity characterized by changes in ID50 and ID95 values, with isolates from several patients (5/15) showing 100-fold increases in ID50. The latter isolates were also insensitive to the related compound, 3'-azido, 3'deoxyuridine (AZdU), but not to the other antiretroviral compounds, dideoxycytidine (ddC), didehydrodideoxythymidine (d4T), carbovir or foscarnet (PFA). With the documentation of this potentially important phenotype and with the availability of methods to make additional isolates from other patients and to assay reliably for drug sensitivity, a number of research objectives can be pursued. We plan: Specific Aim 1: To determine the mechanism of drug resistance. Specific Aim 2: To conduct genetic characterization of the resistance mutation(s). Specific Aim 3: To determine the mechanism by which phosphatidylAZT maintains its activity against AZT resistant isolates of HIV. Specific Aim 4: To determine the virulence and in vivo susceptibility to AZT of the sensitive/resistant isolates of HIV. Specific Aim 5: To determine the frequency of emergence of AZT resistant mutants at different stages of disease. Specific Aim 6: To determine the frequency of emergence of resistant isolates during therapy with drugs other than AZT.

在过去的一年中，我们从Over获得了60多个HIV-1 40名患者，其中许多患者一直在接受AZT长期治疗（3'-- 方济多，3'-脱氧胸苷，Zidovudine）用于艾滋病或与艾滋病有关的高级艾滋病 复杂的。 我们还在CD4-Hela中开发了减少斑块测定法 允许可重现药物定量测定的细胞系 这些分离株的敏感性。 50％的抑制（ID50）值 来自0.01MUM和 0.05MUM。 相比之下，大多数来自接受过AZT的患者的分离株 六个月或更长时间表现出降低的灵敏度，其特征是 ID50和ID95值的变化，来自几名患者的分离株 （5/15）ID50显示100倍。 后一个分离株也是 对相关化合物，3'-齐多，3'Deoxyuridine（azdu）不敏感，但是 不适合其他抗逆转录病毒化合物，二氧辛丁丁（DDC）， 二羟基甲酰胺氨基（D4T），Carbovir或Foscarnet（PFA）。 与 这种潜在重要表型的文档以及 从其他患者中产生其他分离株的方法的可用性 并可靠地分析药物敏感性，许多研究 可以追求目标。 我们计划： 特定目的1：确定耐药性机理。 具体的 目标2：进行电阻的遗传表征 突变。 特定目的3：确定该机制 磷脂酰氮对AZT抗性分离株的活性 艾滋病病毒。 特定目的4：确定毒力和体内 对HIV敏感/抗性分离株的AZT的敏感性。 特定目标5：确定抗AZT的出现频率 在不同疾病阶段的突变体。 特定目标6：确定 在治疗过程中抗性分离株出现的频率 AZT以外的其他药物。