IGF::OT::IGF THE MICROVOA: ENHANCING THE VIRAL OUTGROWTH ASSAY (VOA) WITH MICROTECHNOLOGY

在评估治愈HIV感染的策略时，需要克服的最重要的障碍之一是缺乏一种简单的方法来量化接受高效抗逆转录病毒治疗的个体的静息CD4+记忆T细胞中具有复制能力的HIV潜伏库的大小变化。这些细胞中的大多数HIV DNA代表有缺陷的病毒；只有不到0.01%的高度纯化的静止CD4细胞携带有复制能力的原病毒。因此，基于pcr的方法往往高估了病毒库的大小，并且在病毒生长试验中与产生功能性病毒的细胞数量无关。然而，病毒生长测定是劳动密集型的，需要大量的血液。该项目的目标是设计一个高通量分析平台，可用于可重复地量化从高效抗逆转录病毒治疗个体分离的静息CD4+记忆T细胞中复制能力潜伏HIV库的大小变化。

One of the most significant hurdles to overcome in evaluating strategies to cure HIV infection is the lack of a simple method for quantifying changes in the size of the latent reservoir of replication-competent HIV in resting CD4+ memory T cells in individuals on highly effective antiretroviral therapy. Most of the HIV DNA in these cells represents defective virus; less than 0.01% of highly purified resting CD4 cells harbor replication-competent provirus. As a result, PCR-based methods tend to over-estimate the size of the reservoir and do not correlate with the number of cells producing functional virus in a viral outgrowth assay. However, viral outgrowth assays are labor-intensive and require large volumes of blood. The goal of this project is to design a high-throughput assay platform that can be used to reproducibly quantify changes in the size of the replication-competent latent HIV reservoir in resting CD4+ memory T cells isolated from individuals on highly effective antiretroviral therapy.