Specific killing of latently HIV-1-infected cells after provirus reactivation

原病毒重新激活后特异性杀死潜伏 HIV-1 感染的细胞

• 批准号: 8462367
• 负责人: Qigui Q Yu
• 金额: $ 19.5万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-01 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8462367
• 关键词: Anti-Retroviral Agents; Antibodies; Antibody Formation; Autologous; Biological Assay; CD4 Positive T Lymphocytes; Cell Line; Cells; Complement; Complement Activation; Cytolysis; Detection; Diagnostic; Failure; Gene Expression; Goals; Grant; HIV-1; Highly Active Antiretroviral Therapy; Human; Immune response; Immunity; In Vitro; Individual; Infection; Interleukin-7; Life; Measurable; Mediating; Medical; Memory; Modeling; Morbidity - disease rate; Patients; Pharmaceutical Preparations; Phase; Production; Proliferating; Proviruses; Reporting; Research; Residual state; Rest; Serum; Surface; T-Lymphocyte; Testing; Vaccines; Valproic Acid; Viral; Viral Cytopathogenic Effect; Viral Proteins; Viremia; Virion; Virus; Vorinostat; antiretroviral therapy; cell type; effective therapy; experience; global health; improved; in vivo; inhibitor/antagonist; innovation; killings; member; mortality; novel strategies; pandemic disease; prostratin; public health relevance; purge

DESCRIPTION (provided by applicant): The HIV-1 pandemic has claimed over 20 million lives, with 38.6 million people worldwide currently infected, and will continue to be a significant global health problem as there is no vaccine available. Currently, the only effective treatment available to HIV-1 infection is HAART (highly active antiretroviral therapy), which has led to a profound reduction in HIV-1-related morbidity and mortality. However, HAART fails to eliminate the virus in vivo, mainly due to the persistent existence of long-lived latently-infected cells harboring replication-competent proviruses. Efforts to purge these infected cells have focused on reactivation of the proviruses. It is presumed that these infected cells will be killed after reactivation of virus gene expression by viral cytopathic effects (CPEs), host immune responses or both. Several stimulants including IL-7, valproic acid (VPA), suberoylanilide hydroxamic acid (SAHA) and prostratin have been explored to force activation of proviruses in latently-infected resting CD4+ T cells that constitute the major reservoir of HIV-1 in vivo. However, treatment with IL-7 or VPA in patients on HAART has failed to reduce HIV-1 latency, suggesting that these agents alone are not sufficient to induce killing of latently-infected cells. We propose to develop an "activation-killing" approach by combining provirus stimulants with an agent that is able to allow anti-HIV-1 immunity naturally mounted in patients during HIV-1 infection to specifically kill latently-infected cells after provirus reactivation. Studies have shown that both HIV-1-infected cells and virions use their surface regulators of complement activation (RCA) to resist antibody- dependent complement-mediated lysis (ADCML), which explains why vigorous and sustained anti-HIV-1 envelope (Env) antibody (Ab) responses in almost all infected individuals fail to control HIV-1 infection. We and others have reported that blocking human CD59 (hCD59), a key member of RCA, renders both HIV-1-infected cells and virions sensitive to ADCML. We therefore hypothesize that addition of an hCD59 inhibitor to SAHA, prostratin or both will enable anti-HIV-1 Env Abs naturally present in HIV-1-infected individuals to trigger ADCML of latently-infected T cells after provirus reactivation. The provirus stimulants will activate proviruses to express viral proteins on the surface of latently-infected cells that will be killed by ADCML if hCD59 is inhibited. We will use the human T cell line ACH2, a well-characterized model of HIV-1-latency, to develop such an approach (Aim 1, R21 Phase). We will verify whether the approach is able to trigger ADCML of the primary CD4+ T cells latently infected with HIV-1 in vitro (Aim 2, R33 Phase). We will then determine if this approach can allow anti-HIV-1 Abs naturally present in the sera of infected patients to trigger autologous ADCML of their own latently-infected resting CD4+ T cells (Aim 3, R33 Phase). Our project is innovative and significant because we aim to develop a novel approach for purging the persistent reservoir of HIV-1 in patients on HAART. This approach may simultaneously target various latently infected cells and residual viremia, potentially leading to a broader impact and improved efficacy against the various persistent HIV-1 reservoirs.

描述（由申请人提供）：HIV-1 大流行已夺去了超过 2000 万人的生命，目前全世界有 3860 万人受到感染，并且将继续成为一个重大问题。 由于没有可用的疫苗，全球健康问题。目前，针对 HIV-1 感染唯一有效的治疗方法是 HAART（高效抗逆转录病毒疗法），它已大大降低了 HIV-1 相关的发病率和死亡率。然而，HAART未能在体内消灭病毒，这主要是由于携带具有复制能力的原病毒的长寿命潜伏感染细胞的持续存在。清除这些受感染细胞的努力集中在原病毒的重新激活上。据推测，在病毒细胞病变效应（CPE）、宿主免疫反应或两者共同作用重新激活病毒基因表达后，这些受感染的细胞将被杀死。包括 IL-7、丙戊酸 (VPA)、辛二酰苯胺异羟肟酸 (SAHA) 和 Prostratin 在内的几种兴奋剂已被探索用于强制激活潜伏感染的静息 CD4+ T 细胞中的原病毒，这些细胞构成体内 HIV-1 的主要储存库。然而，接受HAART的患者接受IL-7或VPA治疗未能减少HIV-1潜伏期，这表明单独使用这些药物不足以诱导杀死潜伏感染细胞。我们建议开发 一种“激活杀伤”方法，将原病毒兴奋剂与一种能够使 HIV-1 感染期间患者体内自然产生的抗 HIV-1 免疫力特异性杀死的药物相结合 原病毒重新激活后潜伏感染的细胞。研究表明，HIV-1 感染的细胞和病毒体均利用其表面补体激活调节因子 (RCA) 来抵抗抗体依赖性补体介导的裂解 (ADCML)，这解释了为什么几乎所有感染个体中强烈且持续的抗 HIV-1 包膜 (Env) 抗体 (Ab) 反应无法控制 HIV-1 感染。我们和其他人已经报道，阻断人类 CD59 (hCD59)（RCA 的关键成员）会使 HIV-1 感染的细胞和病毒颗粒对 ADCML 敏感。因此，我们假设在 SAHA、前列腺素或两者中添加 hCD59 抑制剂将使 HIV-1 感染个体中天然存在的抗 HIV-1 Env Ab 能够在原病毒重新激活后触发潜伏感染 T 细胞的 ADCML。原病毒刺激剂将激活原病毒，在潜伏感染的细胞表面表达病毒蛋白，如果 hCD59 受到抑制，这些细胞将被 ADCML 杀死。我们将使用人类 T 细胞系 ACH2（一种经过充分表征的 HIV-1 潜伏期模型）来开发这种方法（目标 1，R21 阶段）。我们将验证该方法是否能够在体外触发潜伏感染 HIV-1 的原代 CD4+ T 细胞的 ADCML（目标 2，R33 阶段）。然后，我们将确定这种方法是否可以允许感染患者血清中天然存在的抗 HIV-1 抗体触发其自身潜伏感染的静息 CD4+ T 细胞的自体 ADCML（目标 3，R33 阶段）。我们的项目具有创新性且意义重大，因为我们的目标是开发一种新方法来清除接受 HAART 治疗的患者体内持续存在的 HIV-1 病毒库。这种方法可以同时针对各种潜伏感染细胞和残留病毒血症，从而可能产生更广泛的影响并提高针对各种持续性 HIV-1 病毒库的功效。