HIV CORECEPTOR SHIFT

描述(由申请人提供)：大约一半的HIV感染者会发生辅受体移位，并通过使用CCR5拮抗剂治疗来促进。这项拟议的工作使用了一组独特的CCR5/CXCR4嵌合辅助受体和一组辅助受体移位HIV-1分离株来检查HIV进入所需的CXCR4的临界域。该方法包括对来自U87的准种HIV-1 gp120序列进行超高密度序列分析。目的1.HIV-1 gp120序列中与VCVr耐药有关的比例是多少？VCVs和VCVR分离株的HIV-1 gp120序列有何不同？为此，U87.CD4.R5.X4细胞将在有或没有VCV的情况下用患者特有的病毒文库感染U87.CD4.R5.X4细胞，并对感染的细胞DNA进行超密集序列分析。目的2.HIV-1 gp120序列中使用CXCR4的比例是多少？gp120序列与不使用CXCR4的序列有何不同？为此，U87.CD4.R5和U87.CD4.X4细胞将被患者特异性的病毒文库感染，并对感染的细胞DNA进行超高密度序列分析。目的3.什么是CXCR4用于感染的最小结构域？利用ECL3或ECL2的HIV-1 gp120序列与不利用这些结构域的序列有何不同？为此，表达嵌合CCR5/CXCR4共受体的U87.CD4细胞将被患者特异性的病毒库感染，并对感染的细胞DNA进行超高密度序列分析。有关HIV进入所需的CXCR4最小结构域使用的分子细节将为指导更有效的抗病毒药物的开发提供关键信息。

DESCRIPTION (provided by applicant): Coreceptor shift occurs in approximately half of all HIV infected individuals, and is promoted by use of CCR5 antagonist therapy. The proposed work uses a unique panel of CCR5/CXCR4 chimeric coreceptors and a panel of coreceptor shift HIV-1 isolates to examine the critical domains of CXCR4 required for HIV entry. The methodology includes ultra dense sequence analysis of HIV-1 gp120 sequences of quasispecies from U87.CD4 cells infected with env amplicons from a panel of dual/mixed isolates selected during CCR5 antagonist therapy in patient-specific virus libraries. Aim 1. What proportion of HIV-1 gp120 sequences are responsible for VCVr resistance and how do HIV- 1 gp120 sequences of VCVs and VCVr isolates differ? For this purpose, U87.CD4.R5.X4 cells will be infected with patient-specific virus libraries in the presence or absence of VCV, and infected cell DNA subjected to ultra dense sequence analysis. Aim 2. What proportion of HIV-1 gp120 sequences utilize CXCR4 for entry and how to the gp120 sequences differ from those that can not utilize CXCR4? For this purpose, U87.CD4.R5 and U87.CD4.X4 cells will be infected with patient-specific virus libraries, and infected cell DNA subjected to ultra dense sequence analysis. Aim 3. What are the minimal domains of CXCR4 utilized for infection and how do HIV-1 gp120 sequences utilizing ECL3 or ECL2 differ from those that do not utilize these domains? For this purpose, U87.CD4 cells expressing chimeric CCR5/CXCR4 coreceptors will be infected with patient-specific virus libraries, and infected cell DNA subjected to ultra dense sequence analysis. Molecular details on the use of minimal domains of CXCR4 required for HIV entry will provide critical information to guide the development of more effective antiviral agents.