Optimized Adaptation of Simian-tropic R5 HIV Clade C to Pig-tailed Macaques

猿猴嗜R5 HIV Cclade C对猪尾猕猴的优化适应

• 批准号: 8714894
• 负责人: Ruth Margrit Ruprecht
• 金额: $ 90.38万
• 依托单位: TEXAS BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-15 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8714894
• 关键词: AIDS/HIV problem; Ablation; Acquired Immunodeficiency Syndrome; Acute; Africa South of the Sahara; Animals; Antiviral Agents; Antiviral Therapy; Architecture; Area; Asorbicap; B-Lymphocytes; Biological; Biopsy; Blood Cells; Brain; CD8-Positive T-Lymphocytes; CD8B1 gene; Capsid Proteins; Cells; Cerebrospinal Fluid; Chimera organism; Consequences of HIV; Cytidine Deaminase; Developing Countries; Development; Disease Progression; Drug usage; Epidemic; Evolution; Family suidae; Generations; Genetic; HIV; HIV Infections; HIV-1; HIV-2; Host Defense; Human; Immunity; Immunocompetent; In Vitro; India; Infection; Link; Long Terminal Repeats; Lymphocyte Depletion; MS4A1 gene; Macaca; Macaca mulatta; Macaca nemestrina; Modality; Modeling; Molecular; Molecular Cloning; Molecular Evolution; Mutate; Pan Genus; Pathogenicity; Peptide Hydrolases; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Phase; Pilot Projects; Plasma; Play; Prevalence; Prevention; Prevention strategy; Primate Lentiviruses; Primates; Property; Protein Isoforms; Proteins; Protocols documentation; RNA-Directed DNA Polymerase; Recombinants; Regimen; Role; Serial Passage; Site; Study models; Tail; Testing; Time; Tissues; Titrations; Tropism; Tumor Necrosis Factor-alpha; Vaccines; Variant; Vertebral column; Viral; Viremia; Virion; Virus; Work; Zambia; adaptive immunity; advanced disease; base; blocking factor; cytokine; fitness; improved; in vivo; lymph nodes; microbicide; mutant; nonhuman primate; novel strategies; pre-clinical; rectal; research study; simian human immunodeficiency virus; therapy development; tool; treatment strategy

DESCRIPTION (provided by applicant): HIV clade C (HIV-C) causes >56% of all cases of HIV/AIDS worldwide and predominates in sub-Saharan Africa and India. A non-human primate (NHP) model would greatly benefit the preclinical development of prevention strategies, including drugs, microbicides, and vaccines. Due to host restriction factors, HIV-1 replicates only in humans and chimpanzees, but not in other NHP species. The construction of simian-human immunodeficiency viruses (SHIVs) encoding HIV-1 vpu, tat, rev and env has yielded pathogenic models in rhesus macaques (RMs) or pig-tailed macaques (PMs~ Macaca nemestrina). A primate lentivirus model based mostly upon HIV-1 but replication-competent in macaques would greatly facilitate the preclinical development of new treatment or prevention strategies. Recently, different host restriction factors that block HIV-1 replication in RM cells have been recognized, namely 1) TRIM5?, which targets HIV-1 capsid proteins and blocks early post-entry replication steps, and 2) APOBEC3G (A3G), a cytidine deaminase that is not recognized by HIV-1 Vif and therefore escapes inhibition by this virion-associated protein. Unlike RMs, PMs lack a TRIM5 isoform, which may explain why PMs supported limited acute viremia of unmodified HIV-1 in earlier studies. Replacing HIV-1 vif with that of SIVmac239, SIVmne or HIV-2 has yielded chimeras that replicated for extended periods of time in PMs~ however, these recombinants were based upon the X4-tropic clade B clones, and did not result in persistent viremia and signs of disease progression. We have generated a simian-tropic HIV-C clone, termed stHIV-C, by replacing HIV vif with SIVmac239 vif in an infectious molecular HIV-C clone from Zambia. The chimera is replication-competent in peripheral blood mononuclear cells (PBMC) of M. nemestrina. In a pilot study, the new stHIV-C-vif239 chimera also induced viremia in two PMs~ however, adaptation is incomplete. The Specific Aims are to: 1. Select stHIV-C progeny with improved replication fitness in PMs under prolonged depletion of CD8+ and CD20+ cells~ 2. Further adapt progeny virus by serial passage through immunocompetent PMs, re-isolate virus from the last recipient, generate a large stock, and characterize it in vitro. 3. Perform an intrarectal titration and assess viral pathogenicity 4. Assess the molecular evolution of stHIV-C in the absence and presence of adaptive immunity and during disease progression. We will also compare viral evolution in different compartments (PBMC, lymph nodes, and brain). The proposed experiments are significant because they focus on the development of an NHP model of HIV-C, which predominates in developing countries where the AIDS epidemic continues to escalate and will thus create a new tool for the preclinical development of different strategies for prevention or eradication.

描述（由申请人提供）：HIV分支C （HIV-C）占全球所有HIV/AIDS病例的60%，在撒哈拉以南非洲和印度占主导地位。非人类灵长类动物（NHP）模型将极大地有利于预防策略的临床前开发，包括药物、杀微生物剂和疫苗。由于宿主的限制因素，HIV-1仅在人类和黑猩猩中复制，而在其他NHP物种中不复制。编码HIV-1 vpu， tat， rev和env的猿人免疫缺陷病毒（SHIVs）的构建已经在恒河猴（RMs）或长尾猕猴（PMs~ Macaca nemestrina）中建立了致病模型。灵长类慢病毒模型主要基于HIV-1，但在猕猴中具有复制能力，这将极大地促进新的治疗或预防策略的临床前开发。最近，在RM细胞中发现了阻断HIV-1复制的不同宿主限制因子，即1)TRIM5？2) APOBEC3G (A3G)，一种胞苷脱氨酶，不被HIV-1 Vif识别，因此逃脱了这种病毒粒子相关蛋白的抑制。与RMs不同，pmms缺乏TRIM5亚型，这可能解释了为什么pmms在早期研究中支持有限的未修饰HIV-1急性病毒血症。用SIVmac239取代HIV-1 vif， sivne或HIV-2产生了嵌合体，在pm中复制了很长一段时间~然而，这些重组是基于X4-tropic枝B克隆，不会导致持续的病毒血症和疾病进展的迹象。我们通过用SIVmac239 vif替换来自赞比亚的传染性分子HIV- c克隆中的HIV- c病毒，生成了一个类人猿的HIV- c克隆，称为shiv - c。嵌合体在nemestrina的外周血单核细胞（PBMC）中具有复制能力。在一项初步研究中，新的stHIV-C-vif239嵌合体也在两个pm中诱导了病毒血症~但是，适应是不完整的。具体目标是：1。选择在CD8+和CD20+细胞长时间耗竭的pmms中具有更好复制适应性的stHIV-C后代~ 2。通过具有免疫功能的pmms连续传代，进一步适应子代病毒，从最后一个受体中重新分离病毒，产生大量的病毒，并在体外鉴定。3. 进行直肠内滴注并评估病毒致病性。评估在缺乏和存在适应性免疫以及疾病进展期间stHIV-C的分子进化。我们还将比较病毒在不同区室（PBMC、淋巴结和脑）中的进化。拟议的实验意义重大，因为它们侧重于开发艾滋病毒- c的NHP模型，这种模型在艾滋病流行病继续升级的发展中国家占主导地位，因此将为临床前制定不同的预防或根除战略创造一种新工具。