Determinants of retroviral replication in non-native hosts for modeling HIV infection

用于模拟 HIV 感染的非本地宿主逆转录病毒复制的决定因素

• 批准号: 10655753
• 负责人: Amit Sharma
• 金额: $ 67.97万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-17 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10655753
• 关键词: Affect; Binding; Binding Sites; Biological; Biological Models; CD4 Positive T Lymphocytes; Cells; Data; Development; Genes; Glycoproteins; Goals; HIV Infections; HIV-1; Human; Immune; In Vitro; Infection; Integral Membrane Protein; Integration Host Factors; Interferon Type I; Interferons; Laboratories; Link; Lymphocyte; Macaca; Maps; Modeling; Molecular; Molecular Conformation; Mucous Membrane; Mutation; Pathogenicity; Polysaccharides; Prevention; Primary Infection; Process; Production; Proteins; Research Proposals; Resistance; Role; SIV; Serial Passage; Structure; T-Lymphocyte Subsets; Time; Vaccines; Variant; Viral; Virion; Virus; Virus Diseases; Virus Replication; base; chronic infection; efficacious intervention; improved; in vivo; insight; macrophage; memory CD4 T lymphocyte; monocyte; neutralizing antibody; non-Native; prevent; rational design; response; simian human immunodeficiency virus; transmission process; virus tropism

PROJECT SUMMARY/ABSTRACT HIV-1 does not persistently infect macaques due to restriction by several species-specific host factors necessitating the use of chimeric SIV/HIV-1 viruses (SHIVs) as surrogates to model HIV-1 infection in macaques. Infection of macaques with SHIVs is the most preferred model system for vaccine and prevention studies because SHIVs encode HIV-1 Envelope glycoprotein (Env) – a key target of HIV-1 neutralizing antibodies. Because the goal of vaccines is to prevent new infection, SHIVs based on circulating, transmitted forms of Env variants are desired as challenge viruses. Existing SHIV/macaque models typically employ SHIVs that encode HIV-1 Env from laboratory-adapted viruses, whose neutralization sensitivities differ from circulating Env variants. This significantly limits the ability of the existing SHIV/macaque models to predict efficacious intervention(s) in humans. Development of SHIVs encoding circulating Env variants has been extremely challenging, mainly because such SHIVs replicate poorly in macaques, if at all. To increase their replication and pathogenicity, SHIVs require extensive adaptation in vivo via serial passage in macaques. The process of serial macaque passage results in accumulation of adaptive mutations in Env that facilitates robust replication. Serial passage is typically performed within the first two weeks of infection, a time during which macaques mount a robust type-I interferon (IFN) response to infection. The host IFN response presents an early barrier against infection because production of IFNs upregulates expression of several IFN-stimulated genes (ISGs), which results in induction of an ‘antiviral state’. Proteins encoded by certain ISGs, referred to as restriction factors, act as potent barriers against cross-species lentiviral transmission. Thus, macaque restriction factors have the potential to block SHIV infection as they can antagonize HIV-1 Env. We recently identified macaque interferon-induced transmembrane proteins (IFITMs) as ISGs that selectively restrict replication of SHIVs encoding circulating HIV-1 Env variants. Our preliminary results suggest that unpassaged SHIV is potently inhibited by IFN in macaque lymphocytes. In contrast, serial passaged SHIVs are resistant to IFN. We found that the loss to two N-linked glycans in Env upon serial passage is sufficient to increase replication and confer resistance to IFN. This research proposal will: 1) characterize the adaptive changes in Env of serial passaged SHIVs that increase replication and IFN resistance; 2) determine the role of N-linked Env glycans in SHIV infection of primary macaque immune cell subsets ex vivo, and mucosal transmission and pathogenicity of SHIVs in vivo; and 3) evaluate the contribution of five macaque IFITM homologs, which are upregulated by IFN, in restriction of unpassaged, IFN-sensitive SHIV. Upon completion, this study will provide mechanistic insights at the host-viral interface that drive selection, adaptation, and pathogenicity of SHIVs in macaques.

项目摘要/摘要 HIV-1由于几个规格特定的宿主因子的限制而不会持续感染猕猴 需要将嵌合SIV/HIV-1病毒（SHIV）用作猕猴中HIV-1感染的替代物。 猕猴感染Shivs是疫苗和预防研究的最优选模型系统 因为SHIV编码HIV-1包膜糖蛋白（ENV） - HIV-1中和抗体的关键靶标。 因为疫苗的目的是防止新感染，所以基于循环，传播形式的湿透者 需要变体作为挑战病毒。现有的Shiv/猕猴模型通常采用编码的SHIVS 来自实验室适应的病毒的HIV-1 Env，其神经抑制剂与循环ENV变体不同。 这显着限制了现有的Shiv/猕猴模型预测有效干预措施的能力 人类。编码循环ENV变体的Shivs的开发极为挑战，主要是 因为这样的动摇在猕猴中复制不佳，如果有的话。为了增加其复制和致病性，Shivs 需要通过猕猴的串行通道在体内进行广泛的适应。串行猕猴密码的过程 导致在ENV中积累自适应突变，从而促进了鲁棒的复制。 串行通道通常在感染的前两周内进行，在此期间猕猴 安装对感染的强大I型干扰素（IFN）反应。主持人IFN响应提出了早期的障碍 反对感染，因为产生IFN会上调几个IFN刺激的基因（ISG）的表达， 这导致诱导“抗病毒状态”。由某些ISG编码的蛋白质，称为限制 因素，充当跨物种慢病毒传播的潜在障碍。那是猕猴的限制因素 有可能阻止SHIV感染，因为它们可以拮抗HIV-1 Env。我们最近确定了猕猴 干扰素诱导的跨膜蛋白（IFITM）与ISGs选择性限制了SHIV的复制 编码循环HIV-1 Env变体。 我们的初步结果表明，在猕猴淋巴细胞中IFN可能会抑制未通透的SHIV。 相比之下，串行传递的湿气对IFN具有抗性。我们发现在Env中损失了两个N连接的聚糖 串行时，足以增加对IFN的复制和会议抵抗力。该研究建议将： 1）表征会增加复制和IFN的串行传递SHIV的自适应变化 反抗; 2）确定N连锁的Env Glycans在原发性猕猴免疫细胞的湿腹膜上的作用 子集在体内的体内，粘膜传播和致病性； 3）评估贡献 在未经通道的IFN敏感的限制下，IFN更新的五个猕猴IFITM同源物中 湿婆。完成后，这项研究将在宿主 - 病毒接口上提供机械见解，以驱动选择， 猕猴中湿虎皮的适应性和致病性。