Evaluation of the sensitivity of Ghanaian HIV-1 strains to protease inhibitors

加纳 HIV-1 毒株对蛋白酶抑制剂的敏感性评估

• 批准号: 15590426
• 负责人: TOKUNAGA Kenzo
• 金额: $ 2.3万
• 依托单位: National Institute of Infectious Diseases
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590426/
• 关键词: HIV-1; Ghanaian strains; Subtypes; Protease inhibitors; Phylogenetic tree; Cassette vector; Drug sensitivity; Structural analysis; プロテアーゼ

Now that highly active antiretroviral therapy(HAART) is being initiated on a large scale in West Africa, it remains controversial whether protease inhibitors(PIs), originally designed and tested against human immunodeficiency virus type 1(HTV-1) subtype B, are equally effective against the non-B subtypes that are prevalent in West African countries. In this study, we investigated whether Ghanaian HIV-1 isolates, as representatives of West African isolates, are susceptible to PIs. We first generated an HIV-1 protease cassette vector proviral DNA fragment carrying a luciferase gene, which allows patient-derived HIV-1 proteases to be inserted and to be subjected to both genotypic and phenotypic assays. HIV-1 protease genes derived from 39 treatment-naive Ghanaian patients were used in this experiment as representatives of West African strains. The cloned patient-derived HIV-1 protease genes were first sequenced and then genetically compared. Phenotypic analysis was performed with Ghanaian … More HTV-1 protease chimeric viruses in the presence of 6 different PIs. Structural models of an HIV-1 protease homodimers were constructed by the molecular modeling software. Genetic analysis of cloned patient-derived HIV-1 protease genes indicated that most of the Ghanaian HIV-1 proteases are placed as subtype CRF02_AG strains, which are phylogenetically distant from subtype B strains, and that Ghanaian HIV-1 proteases do not harbor known major mutations influencing drug resistance but commonly carry 2-3 minor mutations. Phenotypic analysis performed with HIV-1 protease-recombinant viruses in the presence of 6 different PIs revealed that Ghanaian HIV-1 proteases are differentially less susceptible to the PIs. In support of this finding of differential susceptibility, structural analysis showed a significant distortion of nelfinavir, but not of amprenavir, in the Ghanaian protease pocket, suggesting nelfinavir might be less insertable into the Ghanaian protease than into the protease of subtype B. These findings provide implications for the combination of PIs during the introduction of HAART into West Africa. Less

由于高效抗逆转录病毒疗法(HAART)正在西非大规模启动，最初设计和测试的针对人类免疫缺陷病毒1型(HTV-1)B亚型的蛋白酶抑制剂(PI)是否对西非国家流行的非B亚型同样有效，仍存在争议。在这项研究中，我们调查了作为西非分离株代表的加纳HIV-1分离株是否对PI易感。我们首先产生了一个携带荧光素酶基因的HIV-1蛋白酶盒载体前病毒DNA片段，它允许插入患者来源的HIV-1蛋白酶，并接受基因和表型分析。这项实验中使用了来自39名未接受治疗的加纳患者的HIV-1蛋白酶基因作为西非毒株的代表。首先对克隆的患者来源的HIV-1蛋白酶基因进行测序，然后进行遗传比较。用加纳…进行表型分析在6种不同的PI存在时，HTV-1蛋白酶嵌合病毒的数量更多。利用分子建模软件构建了HIV-1蛋白同源二聚体的结构模型。对克隆的患者来源的HIV-1蛋白酶基因的遗传分析表明，加纳的大多数HIV-1蛋白酶属于CRF02_AG亚型，与B亚型毒株的系统发育距离较远，加纳HIV-1蛋白酶没有已知的影响耐药性的主要突变，但通常携带2-3个微小突变。在6种不同PI存在的情况下，对HIV-1蛋白酶重组病毒进行的表型分析表明，加纳的HIV-1蛋白酶对PI的敏感性不同。为了支持这一差异易感性的发现，结构分析显示，加纳蛋白酶口袋中的奈非那韦发生了显著的扭曲，但氨丙那韦没有发生扭曲，这表明奈非那韦可能比B亚型的蛋白酶更难插入加纳的蛋白酶。这些发现为将HAART引入西非期间的PI组合提供了暗示。较少

项目成果

Zheng, Y.-H., Irwin, D., Kurosu, T., Tokunaga, K., Sata, T., Peterlin, B.M.: "Human APOBEC3F is another host factor that blocks HIV-1 replication."J.Virol.. In press. (2004)

Cheng, Y.-H.、Irwin, D.、Kurosu, T.、Tokunaga, K.、Sata, T.、Peterlin, B.M.：“人类 APOBEC3F 是另一种阻断 HIV-1 复制的宿主因子。”J.Virol

Interleukin-8 cross-desensitizes cellular responses to both CCR5 and CXCR4 but inhibits HIV-1 infection only to CCR5 : role of receptor cross-internalization and signal strength.

Interleukin-8 使细胞对 CCR5 和 CXCR4 的反应脱敏，但仅抑制 CCR5 的 HIV-1 感染：受体交叉内化和信号强度的作用。

• 发表时间: 2003
• 期刊: J.Biol.Chem. 278
• 作者: Richardson;R.M.;Tokunaga;K.;Marjoram;R.;Sata;T.;Snyderman;R.
• 通讯作者: R.

Ueno, T., Tokunaga, K., Sawa, H., Maeda, M., Chiba, J., Kojima, A., Hasegawa, H., Shoya, Y., Sata, T., Kurata, T., Takahashi, H.: "Nucleolin, and the packaging signal, Ψ, promote the budding of human immunodeficiency virus type 1(HIV-1)"Microbiol.Immunol.

上野 T.、德永 K.、泽 H.、前田 M.、千叶 J.、小岛 A.、长谷川 H.、翔也 Y.、佐田 T.、仓田 T.、 Takahashi, H.：“核仁素和包装信号 Ψ，促进人类免疫缺陷病毒 1 型 (HIV-1) 的出芽”Microbiol.Immunol。

Human APOBEC3F is another host factor that blocks human immunodeficiency virus type 1 replication

• DOI: 10.1128/jvi.78.11.6073-6076.2004
• 发表时间: 2004-06-01
• 期刊: JOURNAL OF VIROLOGY
• 影响因子: 5.4
• 作者: Zheng, YH;Irwin, D;Peterlin, BM
• 通讯作者: Peterlin, BM

Human topoisomerase I promotes HIV-1 proviral DNA synthesis : implications for the species specificity of HIV-1 infection.

人类拓扑异构酶 I 促进 HIV-1 前病毒 DNA 合成：对 HIV-1 感染的物种特异性的影响。

• 发表时间: 2003
• 期刊: Proc.Natl.Acad.Sci.U.S.A. 100
• 作者: Shoya;Y.;Tokunaga;K.;Sawa;H.;Maeda;M.;Ueno;T.;Yoshikawa;T.;Sata;T.;Kurata;T.;Cullen;B.R.;Takahashi;H.
• 通讯作者: H.