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/总结： 常用的HIV-1动物模型包括用猿猴免疫缺陷病毒感染猕猴 (SIV)或含有HIV包膜（Env）或逆转录酶的猿猴-人类免疫缺陷病毒（SHIV）。 这些动物模型在理解HIV发病机制和疾病进展方面非常有用， 以及了解疫苗和药物的功效。然而，HIV-1和HIV-2之间的基因差异 和SIV，以及SHIV中其他HIV-1基因如gag、pol、vif、vpr和nef的缺失限制了 疫苗研究中的这些模型。理想地，HIV-1感染/AIDS的良好动物模型将是 携带HIV-1的猕猴然而，由于逆转录病毒的存在，HIV-1不会在猕猴细胞中复制 制约因素HIV-1可以通过用SIV基因取代其辅助基因来复制， VIF、Vpx、Vpr和Nef，它们可以抵消猕猴细胞中干扰素诱导的限制因子。我确 我以前曾报道过，人-猴免疫缺陷病毒是由HIV-1 NL 4 -3 vif的取代产生的， 用SIV替换的HSIV-vifNL 4 -3可以在猪尾猕猴（PTM）中持续复制。然而，在这方面， 感染并不导致SIV或猿猴-人试验中观察到的高峰病毒血症和设定点病毒载量 猕猴的免疫缺陷病毒（SHIV）感染。为了进一步适应HSIV，我们在体内进行了连续的 传代以增强感染力或复制能力。我们将受感染的血液进行动物间转移 在3种免疫活性PTM中，起始初始接种物含有CXCR 4-（HSIV-vifNL 4 -3 回收自先前感染的猕猴）和嗜CCR 5的HSIV（基于pNL-AD 8的HSIV-vif衍生物 和Bru-Yu 2）。有趣的是，所有的猕猴都表现出接近或高于105拷贝/ml的峰值病毒血症， 复制持续了20多周。我们已经发现了三个嗜CXCR 4的感染性分子 来自第3代猕猴（HSIV-P3 IMC）的克隆（IMC），在整个基因组中具有感兴趣的突变， 也许暗示着对PTM的适应。我们假设HSIV-P3 IMCs的进一步体内传代将导致HSIV-P3 IMCs的进一步增殖。 产生具有增强的复制能力的致病性变体。我们建议在体内进行一系列 在较老的PTM中传代，与幼年/年轻猕猴相比，这可能支持更好的病毒复制。 由于我们仅回收了CXCR 4-嗜性HSIV，我们还建议使用HSIV-P3 IMC作为骨架来开发 CCR 5-嗜性HSIV。这项研究的结果将为生物学的发展提供有价值的见解。 HIV-1感染的相关动物模型，用于疫苗预防HIV传播的临床前评价。