Pigtail Macaque Model of Human-Simian Immunodeficiency Virus Infection

人猿免疫缺陷病毒感染的尾猴模型

• 批准号: 10484431
• 负责人: Rajesh Thippeshappa
• 金额: $ 37.8万
• 依托单位: TEXAS BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-12 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10484431
• 关键词: AIDS prevention; Acquired Immunodeficiency Syndrome; Adolescent; Animal Model; Animals; Anti-Retroviral Agents; Antibody Response; Autopsy; Blood; CCR5 gene; CD4 Positive T Lymphocytes; CXCR4 gene; Cell Count; Cells; Coculture Techniques; DNA; Development; Disease; Disease Progression; Evolution; Generations; Genes; Genetic; Genome; Goals; HIV; HIV-1; Human; Immunocompetent; In Vitro; Infection; Interferon-alpha; Interferons; Macaca; Macaca nemestrina; Measures; Messenger RNA; Modeling; Molecular Cloning; Mutation; Names; Open Reading Frames; Pathogenesis; Pathogenicity; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Plasma; Play; Protein Family; Proteins; RNA Splicing; RNA-Directed DNA Polymerase; Reporting; Research Project Grants; Resistance; Resources; SIV; Site; Time; Tissues; Variant; Vertebral column; Viral; Viral Load result; Viral Proteins; Viremia; Virus; Virus Diseases; Virus Replication; base; efficacy study; experimental study; human model; in vivo; insight; nonhuman primate; novel therapeutics; preclinical evaluation; prevent; simian human immunodeficiency virus; therapeutic vaccine; transmission process; vaccine efficacy; vaccine evaluation; vaccine trial; viral RNA

Abstract/Summary: Commonly used animal models of HIV-1 include infection of macaques with Simian Immunodeficiency Virus (SIV) or Simian-Human Immunodeficiency Virus (SHIV) containing HIV envelope (Env) or reverse transcriptase. These animal models have been extremely useful in understanding HIV pathogenesis and disease progression, as well as understanding the efficacy of vaccines and drugs. However, the genetic difference between HIV-1 and SIV, and the absence of other HIV-1 genes such as gag, pol, vif, vpr, and nef in SHIV limits the utility of these models in vaccine studies. Ideally, good animal model of HIV-1 infection/AIDS would be infection of macaques with HIV-1. However, HIV-1 does not replicate in macaque cells due to the presence of retroviral restriction factors. HIV-1 can be made to replicate by substituting its accessory genes with SIV genes such as vif, vpx, vpr, and nef, which can counteract interferon-induced restriction factors in macaque cells. Indeed, we have previously reported that Human-Simian Immunodeficiency Virus generated by substitution of HIV-1NL4-3 vif with SIV substitution (named HSIV-vifNL4-3) can replicate persistently in pigtail macaques (PTMs). However, infection did not result in high peak viremia and setpoint viral loads as observed during SIV or Simian-Human Immunodeficiency virus (SHIV) infection of macaques. To further adapt HSIV, we performed serial in vivo passaging to enhance infectivity or replicative capacity. We conducted animal-to-animal transfer of infected blood in 3 immunocompetent PTMs with starting initial inoculum containing a mixture of CXCR4- (HSIV-vifNL4-3 recovered from previously infected macaque) and CCR5-tropic HSIV (HSIV-vif derivative based on pNL-AD8 and Bru-Yu2). Interestingly, all the macaques showed peak viremia close to or above 105 copies/ml and virus replication persisted for more than 20 weeks. We have recovered three CXCR4-tropic infectious molecular clones (IMCs) from passage 3 macaque (HSIV-P3 IMCs) with interesting mutations throughout the genome, perhaps suggesting adaptation to PTMs. We hypothesize that further in vivo passaging of HSIV-P3 IMCs will generate pathogenic variants with enhanced replication capacity. We propose to conduct serial in vivo passaging in older PTMs, which may support better virus replication compared to juvenile/younger macaques. Since we recovered only CXCR4-tropic HSIV, we also propose to use HSIV-P3 IMCs as backbones to develop CCR5-tropic HSIV. The results from this study will provide valuable insights into development of biologically relevant animal model of HIV-1 infection for preclinical evaluation of vaccine prevention of HIV transmission.

摘要/摘要： HIV-1的常用动物模型包括猕猴患有猿猴免疫缺陷病毒 （SIV）或含有含HIV包膜（ENV）或逆转录酶的猿猴免疫缺陷病毒（SHIV）。 这些动物模型在理解HIV发病机理和疾病进展方面非常有用， 以及了解疫苗和药物的功效。但是，HIV-1之间的遗传差异 和SIV，以及其他HIV-1基因（例如SHIV中的GAG，POL，VIF，VPR和NEF）等其他HIV-1基因限制了 这些模型在疫苗研究中。理想情况下，HIV-1感染/艾滋病的良好动物模型将是感染 带有HIV-1的猕猴。但是，由于存在逆转录病毒，HIV-1不会在猕猴细胞中复制 限制因素。可以通过用SIV基因（例如 VIF，VPX，VPR和NEF，可以抵消干扰素诱导的猕猴细胞中的限制因子。确实，我们 以前已经报道过，由HIV-1NL4-3 VIF取代产生的人类西米亚免疫缺陷病毒 使用SIV替代（命名为HSIV-VIFNL4-3）可以在尾型猕猴（PTMS）中持续复制。然而， 感染不会导致高峰值病毒血症和固定点病毒载量，如SIV或Simian-Human期间所观察到的 猕猴的免疫缺陷病毒（SHIV）感染。为了进一步调整HSIV，我们在体内进行了串行 传播以增强感染力或复制能力。我们进行了动物到动物的感染血液转移 在3个免疫能力的PTM中，开始含有CXCR4-混合物的初始接种物（HSIV-VIFNL4-3） 从先前感染的猕猴）和CCR5-循环HSIV（基于PNL-AD8的HSIV-VIF衍生物 和bru-yu2）。有趣的是，所有猕猴显示峰值病毒血症接近或以上105份/ml和病毒 复制持续了20周以上。我们恢复了三个CXCR4 - 循环传染性分子 来自第3猕猴（HSIV-P3 IMC）的克隆（IMC），整个基因组中具有有趣的突变， 也许暗示适应PTMS。我们假设HSIV-P3 IMC的进一步传播将 产生具有增强复制能力的致病变体。我们建议在体内进行串行 与少年/年轻的猕猴相比，较旧的PTM的传播可能支持更好的病毒复制。 由于我们仅恢复了CXCR4-Tropic HSIV，因此我们还建议使用HSIV-P3 IMC作为骨架来开发 CCR5 - 热带HSIV。这项研究的结果将为生物学发展提供宝贵的见解 HIV-1感染的相关动物模型，用于预防HIV传播疫苗的临床前评估。