Studies of HIV Latency in Primary CD4 T-Lymphocytes

原代 CD4 T 淋巴细胞中 HIV 潜伏期的研究

• 批准号: 7230693
• 负责人: Miles W. Cloyd
• 金额: $ 11.33万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-30 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7230693
• 关键词: T lymphocyte; culture; memory

DESCRIPTION (provided by applicant): The ability of HIV to reduce its replication and to also go into latency during chronic infection is likely one of the important mechanisms establishing its persistence in the face of the antiviral immune response that is always induced. Moreover, this aspect of HIV may be the main reason HAART does not cure infections. Accordingly, latently infected cells in patients must be eliminated to eventually effect a cure. However, studies in primary CD4 lymphocytes of the molecular mechanisms involved in regulation of HIV latency have been hindered by the scarcity of these cells in vivo. We have developed a culture system that appears to recapitulate what occurs in vivo to generate latently infected CD4 lymphocytes. In this system, normal activated CD4 lymphocytes are rescued from apoptosis that normally occurs in vitro, and these cells can be maintained viably in a resting memory state for many months. Our preliminary studies show that acutely infected, activated normal human CD4 lymphocytes that are rescued and maintained in this culture system result in 5 to 10% of the cells having latent HIV. Therefore, the purpose of this grant is to fully characterize this system to show whether it truly generates latently infected cells as found in vivo. Aim 1 will optimize the conditions for generating HIV-latently infected normal CD4 lymphocytes using this system. Aim 2 will examine whether they are truly latently infected, and determine some of their characteristics. To our knowledge we have developed the only in vitro culture system to rescue activated normal lymphocytes and to maintain memory cells long term, and this system should facilitate studies of chronic HIV infection in normal CD4 lymphocytes and the eventual delineation of molecular mechanisms of HIV latency and reactivation.

描述（由申请人提供）：HIV在慢性感染期间减少复制和进入潜伏期的能力可能是在面对总是诱导的抗病毒免疫反应时建立其持久性的重要机制之一。此外，艾滋病毒的这一方面可能是HAART无法治愈感染的主要原因。因此，必须清除患者体内潜伏感染的细胞，才能最终实现治愈。然而，在原生CD4淋巴细胞中参与调节HIV潜伏期的分子机制的研究由于体内这些细胞的缺乏而受到阻碍。我们已经开发了一种培养系统，它似乎概括了体内产生潜伏感染的CD4淋巴细胞的过程。在这个系统中，正常活化的CD4淋巴细胞从体外正常发生的凋亡中被拯救出来，这些细胞可以在静息记忆状态下维持数月。我们的初步研究表明，急性感染、激活的正常人类CD4淋巴细胞在这种培养系统中被拯救和维持，导致5%至10%的细胞具有潜伏的HIV。因此，这项拨款的目的是充分表征该系统，以显示它是否真的产生体内发现的潜伏感染细胞。目的1将优化使用该系统产生hiv潜伏感染的正常CD4淋巴细胞的条件。目标2将检查他们是否真的是潜伏感染，并确定他们的一些特征。据我们所知，我们已经开发了唯一的体外培养系统来拯救活化的正常淋巴细胞并长期维持记忆细胞，该系统应该有助于研究正常CD4淋巴细胞的慢性HIV感染，并最终描述HIV潜伏期和再激活的分子机制。