Modeling immune impairments and pathogenesis in novel humanized mice for HBV-HIV co-infection

项目摘要 HBV和HIV-1合并感染是常见的，HIV合并感染可加重病毒性肝炎的进展 并加速肝病进展。阐明这些潜在机制的主要障碍是 常见和严重的疾病，从而开发有效的治疗方法，是缺乏小动物模型， 复制HBV和HIV-1的类人感染。这两种病毒都表现出狭窄的宿主范围， 主要限于人类和其他类人猿。我们的实验室（Ploss和Su）已经开发出新型人源化小鼠 与人肝脏和人免疫细胞共移植的小鼠（FNRGF/A2-hu HEP/HSC小鼠），与人 肝细胞和改善的人髓样细胞/DC和NK细胞亚群，这些亚群在常规免疫中代表性不足。 人源化小鼠模型。我们已经证明，这些改进产生疫苗诱导的免疫原性 类似于已经接种相同的明确定义的YFV-17 D疫苗的人的特征。我们 我们将利用这些有希望的突破来研究以下内容：1）我们将感染FNRGF/A2的队列- 用人HLA匹配的肝细胞和携带HBV的人造血细胞双重移植的hu小鼠和/或 HIV-1我们将纵向监测和量化HBV和HIV病毒血症和免疫激活，包括抗原- 特异性CD 8 + T细胞和肝脏疾病进展（目的1A）。我们还将在HBV/HIV队列中进行分析 已经用HAART（包括HBV聚合酶抑制剂）治疗以抑制HIV-1的共感染小鼠 和HBV，以准确模拟临床相关情况（目的1B）。2)我们将研究HIV/HBV诱导的T细胞 损伤我们将监测HIV和HBV特异性T细胞上的T细胞耗竭标志物，并分析功能性T细胞的功能。 用单细胞转录组学研究HBV/HIV合并感染时抗原特异性CD 8 + T细胞的损伤 2A）。我们还将评估阻断INFAR 1对HIV/HBV诱导的免疫损伤的影响。我们会监察 逆转HIV和HBV特异性T细胞的T细胞耗竭功能和IFN诱导的功能谱 HBV/HIV共感染期间抗原特异性CD 8 + T细胞的损伤（目的2B）。3)我们将调查如何 HIV-1感染使体内人源化小鼠肝脏中HBV诱导的致病性巨噬细胞升高（目的3A）。 我们还将研究HIV和HBV如何与M2样巨噬细胞相互作用以激活人类星状细胞（Aim 3B）。 最后，我们将研究抑制M2样巨噬细胞如何预防或逆转HIV/HBV诱导的肝脏疾病， 人源化小鼠（Aim 3C）。 因此，我们将利用我们在HIV-1/HBV（Su）病毒学和免疫学方面的广泛互补专业知识， 肝炎病毒（Ploss）和人源化小鼠技术来实现令人兴奋的目标。我们的工作将向前推进 HBV和HIV-1研究领域，表明一种新的小动物模型可以成功地用于 了解HIV/HBV合并感染和免疫反应，并为相关肝脏疾病的治疗建模。

Project Summary Co-infection with HBV and HIV-1 is common, and HIV co-infection can exacerbate progression of viral hepatitis and accelerate liver disease progression. A major hurdle to elucidating potential mechanisms underlying these common and serious diseases, and thus to developing effective therapies, is the lack of small animal models that reproduce human-like infection with HBV and HIV-1. Both viruses exhibit a narrow host range in which infection is largely limited to humans and other great apes. Our labs (Ploss and Su) have developed novel humanized mice that are co-engrafted with human livers and human immune cells (FNRGF/A2-hu HEP/HSC mice), with human hepatocytes and improved human myeloid cell/DC and NK cell subsets that are underrepresented in conventional humanized mouse models. We have demonstrated that these refinements yield vaccine-induced immunogenicity profiles resembling those of humans who have been vaccinated with the same well-defined YFV-17D vaccine. We will capitalize on these promising breakthroughs to investigate the following: 1) We will infect cohorts of FNRGF/A2- hu mice dually engrafted with human HLA-matched hepatocytes and human hematopoietic cells with HBV and/or HIV-1. We will monitor, and quantify longitudinally, HBV and HIV viremia and immune activation, including antigen- specific CD8+ T cells and liver disease progression (Aim 1A). We will also perform analyses in cohorts of HBV/HIV co-infected mice which have been treated with HAART (including HBV polymerase inhibitors), to suppress HIV-1 and HBV, to model accurately clinically relevant situations (Aim 1B). 2) We will study HIV/HBV-induced T cell impairment. We will monitor T cell exhaustion markers on HIV and HBV specific T cells and profile functional impairments in antigen specific CD8+ T cells during HBV/HIV co-infections using single-cell transcriptomics (Aim 2A). We will also evaluate the impact of blocking INFAR1 on HIV/HBV-induced immune impairment. We will monitor reversion of T cell exhaustion functions of HIV- and HBV-specific T cells and profile IFN-induced functional impairments in antigen specific CD8+ T cells during HBV/HIV co-infections (Aim 2B). 3) We will investigate how HIV-1 infection elevates HBV-induced pathogenic macrophages in the liver of humanized mice in vivo (Aim 3A). We will also study how HIV and HBV interact with M2-like macrophages to activate human stellate cells (Aim 3B). Finally, we will study how inhibiting M2-like macrophages prevents or reverses HIV/HBV-induced liver diseases in humanized mice (Aim 3C). We will thus capitalize on our extensive complimentary expertise in virology and immunology of HIV-1/HBV (Su), hepatitis viruses (Ploss) and in humanized mouse technology to achieve the exciting aims. Our work will advance the field of HBV and HIV-1 research by showing that a novel small animal model can be successfully used to understand HIV/HBV coinfection and immune responses, and to model treatments for the associated liver diseases.