A mouse viral outgrowth assay for the detection of residual HIV-1 reservoirs

用于检测残留 HIV-1 病毒库的小鼠病毒生长测定

• 批准号: 8965588
• 负责人: JOEL N BLANKSON
• 金额: $ 40.5万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-08 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8965588
• 关键词: A Mouse; Adoptive Transfer; Allogeneic Bone Marrow Transplantation; Allogenic; Antigens; B-Lymphocytes; Berlin; Biological Assay; Bone Marrow; Bone Marrow Transplantation; Boston; CCR5 gene; CD28 gene; CD3 Antigens; CD4 Positive T Lymphocytes; Case Study; Cell Death; Cells; DNA; Data; Defective Viruses; Detection; Development; Flushing; Frequencies; Future; Gut associated lymphoid tissue; HIV; HIV-1; Hematologic Neoplasms; Hematopoietic; Highly Active Antiretroviral Therapy; Human; Immune response; Immunodeficient Mouse; Individual; Infection; Latent Virus; Lead; Malignant Neoplasms; Measures; Mediating; Methods; Mus; Mutation; Natural Killer Cells; Outcome; Patients; Peripheral; Peripheral Blood Mononuclear Cell; Procedures; RNA; Regimen; Reporting; Residual state; Rest; Shock; Stem cell transplant; Study Subject; T-Lymphocyte; Testing; Therapeutic Intervention; Viral; Viral Antigens; Viral Cytopathogenic Effect; Viremia; Virus; Work; antiretroviral therapy; base; experience; immune activation; in vivo; killings; novel; prevent; public health relevance; response; screening

 DESCRIPTION (provided by applicant): Latently infected CD4+ T cells represent the major barrier to HIV-1 eradication. Many strategies are being developed to eliminate these cells, but the field lacks a simple sensitive assay that can screen for these cells. PCR based assays cannot distinguish between replication-competent versus defective virus and recent studies have shown that the current state of the art culture assay measures only a small fraction of the replication-competent virus present. In this proposal we aim to develop a sensitive assay that is capable of selectively amplifying replication-competent virus from latently infected CD4+ T cells. Such an assay could inform the decision of whether or not to discontinue cART in patients after a therapeutic intervention is made. The need for such an assay is highlighted by the recent report of the 2 "Boston patients" who were treated with allogeneic stem cell transplantation for malignancies. No HIV DNA was detected by PCR in either patient from as many as 200 million PBMCs (<0.07 copies/106 PBMCs), but after cART was discontinued there was a rapid rebound in viremia in both patients. This outcome is undesirable is it negates the theoretical advantages to having smaller HIV-1 reservoirs. The development of a simple but sensitive assay that can screen a very large number of CD4+ T cells may help prevent the occurrence of a similar scenario from occurring in the future. This work will be important for the development of strategies for HIV-1 eradication since the assay will help inform the decision of whether or not to discontinue antiretroviral therapy.

 描述(由申请人提供)：潜伏感染的CD4+T细胞是根除HIV-1的主要障碍。人们正在开发许多策略来消除这些细胞，但该领域缺乏一种简单、灵敏的方法来筛选这些细胞。基于聚合酶链式反应的检测无法区分具有复制能力的病毒和缺陷病毒，最近的研究表明，目前最先进的培养检测方法只能测量存在的一小部分具有复制能力的病毒。在这项建议中，我们的目标是开发一种敏感的检测方法，能够选择性地从潜伏感染的CD4+T细胞中扩增具有复制能力的病毒。这样的检测可以为患者在进行治疗干预后是否停止CART的决定提供信息。最近报道的两名“波士顿患者”接受了异基因干细胞移植治疗恶性肿瘤的报道，强调了这种检测的必要性。在多达2亿个PBMC(0.07拷贝/106个PBMC)中，两名患者均未检测到HIV DNA，但在停用CART后，两名患者的病毒血症迅速反弹。这一结果是不可取的，因为它否定了拥有较小的HIV-1宿主的理论优势。开发一种简单但敏感的检测方法，可以筛查大量的CD4+T细胞，可能有助于防止类似情况在未来发生。这项工作对于制定根除HIV-1的战略将是重要的，因为化验将有助于决定是否 停止抗逆转录病毒治疗。