Pigtail Macaque Model of Human-Simian Immunodeficiency Virus Infection

人猿免疫缺陷病毒感染的尾猴模型

基本信息

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
项目类别：
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

项目摘要

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 动物模型包括感染猿猴免疫缺陷病毒的猕猴 (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)。这些动物模型对于了解艾滋病毒发病机制和疾病进展非常有用，以及了解疫苗和药物的功效。然而，HIV-1 之间的基因差异和 SIV，并且 SHIV 中缺乏其他 HIV-1 基因，例如 gag、pol、vif、vpr 和 nef，限制了其实用性疫苗研究中的这些模型。理想情况下，HIV-1 感染/艾滋病的良好动物模型是感染携带 HIV-1 的猕猴。然而，由于逆转录病毒的存在，HIV-1 不能在猕猴细胞中复制。限制因素。 HIV-1可以通过用SIV基因取代其辅助基因来复制，例如 vif、vpx、vpr 和 nef，可以抵消猕猴细胞中干扰素诱导的限制因子。确实，我们此前曾报道，通过替换 HIV-1NL4-3 vif 产生人猿免疫缺陷病毒带有 SIV 替代的病毒（称为 HSIV-vifNL4-3）可以在猪尾猕猴 (PTM) 中持续复制。然而，感染不会导致 SIV 或猿猴实验期间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，我们在体内进行了系列研究传代以增强感染性或复制能力。我们进行了受感染血液的动物间转移在 3 个免疫活性 PTM 中，起始初始接种物含有 CXCR4- (HSIV-vifNL4-3 从先前感染的猕猴中恢复）和 CCR5-tropic HSIV（基于 pNL-AD8 的 HSIV-vif 衍生物）和 Bru-Yu2)。有趣的是，所有猕猴都表现出接近或高于 105 拷贝/毫升的峰值病毒血症，并且病毒复制持续了20多周。我们已经回收了三种CXCR4嗜性感染分子来自第 3 代猕猴 (HSIV-P3 IMC) 的克隆 (IMC)，整个基因组具有有趣的突变，也许建议适应 PTM。我们假设 HSIV-P3 IMC 的进一步体内传代将产生具有增强复制能力的致病变异。我们建议在体内进行系列在较老的 PTM 中进行传代，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 CXCR4-tropic HSIV，因此我们还建议使用 HSIV-P3 IMC 作为骨干来开发 CCR5-tropic 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。