权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Optimization of high throughput viral outgrowth assays for the detection of HIV-1 reservoirs

用于检测 HIV-1 病毒库的高通量病毒生长检测的优化

基本信息

批准号：
10177855
负责人：
JOEL N BLANKSON
金额：
$ 40.94万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-05 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10177855
关键词：
Allogenic Anti-Retroviral Agents Antibodies Berlin Biological Assay Bone Marrow Bone Marrow Transplantation Boston CCR5 gene CD28 gene CD4 Positive T Lymphocytes Case Study Cell Line Cells DNA Defective Viruses Development Frequencies Future Goals HIV HIV Infections HIV-1 Hematologic Neoplasms Immune response Individual Interruption Lead Length Malignant Neoplasms Measurable Measures Methods Mitogens Mus Mutation Outcome Patients Peripheral Blood Mononuclear Cell Pharmaceutical Preparations Primary Infection Procedures Production Proviruses RNA Reporter Genes Reporting Residual state Rest Sampling Shock Stem cell transplant T-Cell Activation T-Lymphocyte Testing Therapeutic Intervention Time Viral Viral Antigens Viral Cytopathogenic Effect Viral reservoir Viremia Virus Virus Latency Virus Replication Work antiretroviral therapy base cohort detection assay experience high throughput screening improved lymphoblast prevent rapid testing viral rebound virus culture

项目摘要

Project Summary 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. We will use the optimized assay we develop to screen CD4+ T cells from patients who were treated with ART during primary infection and correlate the number of latently infected cells as measured in our assay to the time it took for viral rebound to occur when ART was discontinued in these subjects. 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.

项目总结