CNS Myeloid Cells as SIV Reservoirs: Persistent Infection and Rebound

项目摘要(摘要) 防止艾滋病毒在抗逆转录病毒治疗中断(ATI)后反弹仍然是一个主要问题。然而， 事实证明，准确定位病毒反弹的蓄水池细胞是难以捉摸的。一个- 抗逆转录病毒疗法(ART)已经显示出从外周清除生产性感染细胞的有效性， 在大脑方面取得了有限的成功。然而，一旦ART被撤回，病毒就会反弹，脑币中的感染- 伴有神经侵袭和伴随的神经炎症。长期目标是为 识别并防止艾滋病毒在ATI上反弹。本应用程序的目标是开发一种 确定中枢神经系统和其他相关器官中的髓细胞储存库的战略和测量手段。 中心假说是受感染的小胶质细胞和其他髓系细胞将是反弹的来源 ATI之后的病毒。这是基于艺术和艺术跨学科团队之前的工作。 猕猴(Ling)、神经免疫学(MacLean)和HIV-1/SIV分子病毒学(Ling， Maness&Keele)。我们将采用条形码SIV(Keele)、先进的成像技术、细胞培养和Next 条形码区域的生成测序以识别潜伏感染的细胞并确定哪些细胞 ATI上反弹病毒的来源。提出了两个具体目标：具体目标1：确定 在SIV感染的猕猴中，中枢神经系统中的髓细胞含有可重新激活的复制能力病毒 完全压抑的艺术。虽然淋巴细胞在HIV感染中受到的关注最多，但大脑中的髓细胞， 肺和淋巴组织通常被忽视，因为它们是HIV感染的创建者/再创建者的靶细胞。我们的- 这里的假设是，SIV感染的髓系细胞可以被重新激活，从而产生新的感染病毒。 特定目标2：确定中枢神经系统中的髓样细胞是否是ART后反弹病毒的来源 中断(ATI)。我们推测小胶质细胞和其他髓系细胞可能是病毒的来源。 在ATI上。我们关于这一目标的中心假设是大脑特定的条形码和进化的序列将 在ATI后在髓系细胞中鉴定，并与从反弹中分离的序列具有很高的相似性 外周血中的病毒。我们相信，一旦这个项目完成，就有可能确定目标细胞以 在没有继续抗逆转录病毒治疗的情况下防止艾滋病毒反弹。 好了！

Project Summary (Abstract) Preventing rebound of HIV following antiretroviral therapy interruption (ATI) remains a major problem. However, pinpointing the exact cells that serve as reservoirs from which the virus rebounds has proven elusive. An- tiretroviral therapy (ART) has shown effectiveness in removing productively infected cells from the periphery, with limited success in brain. However, once ART is withdrawn, virus rebounds, and infection in the brain coin- cide with neuroinvasion and concomitant neuroinflammation. The Long-Term Goal is to establish a basis for identifying and then preventing HIV rebounding upon ATI. The Objective of this Application is to develop a strategy for identifying, and a means of measuring, the myeloid reservoir in the CNS and other relevant organs. The Central Hypothesis is that infected microglia and other myeloid cells would be the source of rebounding virus following ATI. This is based on previous work from the interdisciplinary team of experience with ART and reservoir research in macaques (Ling), neuroimmunology (MacLean) and HIV-1/SIV molecular virology (Ling, Maness & Keele). We will employ a barcoded SIV (Keele), advanced imaging techniques, cell culture and next generation sequencing of the barcoded region to identify latently infected cells and determine which cells are the source of rebound virus upon ATI. Two Specific Aims are proposed: Specific Aim 1: Determine whether myeloid cells in CNS harbor reactivatable replication-competent virus in SIV-infected macaques under fully suppressive ART. While lymphocytes receive the most attention for HIV infection, myeloid cells in brain, lungs and lymphoid tissues are often overlooked as founder/ refounder target cells of HIV infection. Our hy- pothesis here is that SIV infected myeloid cells can be reactivated to produce infectious virus for new infection. Specific Aim 2: Determine whether myeloid cells in the CNS are a source of rebound virus upon ART interruption (ATI). We hypothesize that microglia and other myeloid cells would be the source of virus re- bound upon ATI. Our central hypothesis for this aim is that brain-specific barcodes and evolved sequences will be identified within myeloid cells following ATI and have high similarity with sequences isolated from rebound virus in peripheral blood. We believe once this project is completed, it will be possible to identify target cells to prevent HIV rebounding in the absence of continued ART. !