HIV CORECEPTOR SHIFT

描述（由申请人提供）：大约一半的HIV感染者发生共受体转移，并通过使用CCR5拮抗剂治疗促进。这项工作使用一组独特的CCR5/CXCR4嵌合共受体和一组共受体移位的HIV-1分离物来检测HIV进入所需的CXCR4的关键结构域。方法包括U87准种HIV-1 gp120序列的超密集序列分析。在患者特异性病毒文库中，在CCR5拮抗剂治疗期间选择的一组双重/混合分离物，感染了env扩增子的CD4细胞。目的1。有多少比例的HIV-1 gp120序列对VCVr的抗性负责？ vcv和VCVr分离株的HIV-1 gp120序列有何不同？为此，请使用U87.CD4.R5。在存在或不存在VCV的情况下，用患者特异性病毒文库感染X4细胞，并对感染细胞的DNA进行超密集序列分析。目标2。有多少比例的HIV-1 gp120序列利用CXCR4进入？ gp120序列与不利用CXCR4的gp120序列有何不同？为此，U87.CD4。R5和U87.CD4。用患者特异性病毒文库感染X4细胞，并对感染细胞DNA进行超密序列分析。目标3。CXCR4用于感染的最小结构域是什么？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，请使用U87。用患者特异性病毒文库感染表达嵌合CCR5/CXCR4共受体的CD4细胞，并对感染细胞DNA进行超密序列分析。关于HIV进入所需的CXCR4最小结构域使用的分子细节将为指导开发更有效的抗病毒药物提供关键信息。

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.