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

项目概要 HBV 和 HIV-1 合并感染很常见，HIV 合并感染会加剧病毒性肝炎的进展 并加速肝脏疾病的进展。阐明这些潜在机制的主要障碍 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 复制类似人类的 HBV 和 HIV-1 感染。这两种病毒都表现出狭窄的宿主范围，感染是 很大程度上仅限于人类和其他类人猿。我们的实验室（Ploss 和 Su）开发了新型人源化小鼠 与人类肝脏和人类免疫细胞（FNRGF/A2-hu HEP/HSC 小鼠）共同移植的 肝细胞和改良的人类骨髓细胞/DC 和 NK 细胞亚群，这些细胞亚群在传统方法中代表性不足 人源化小鼠模型。我们已经证明这些改进可以产生疫苗诱导的免疫原性 其特征与接种过相同明确 YFV-17D 疫苗的人类相似。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- 群体 hu 小鼠双重移植人类 HLA 匹配的肝细胞和带有 HBV 和/或的人类造血细胞 HIV-1。我们将纵向监测和量化 HBV 和 HIV 病毒血症以及免疫激活，包括抗原- 特异性 CD8 T 细胞和肝脏疾病进展（目标 1A）。我们还将对 HBV/HIV 队列进行分析 已接受 HAART（包括 HBV 聚合酶抑制剂）治疗的共感染小鼠，以抑制 HIV-1 和 HBV，以准确地模拟临床相关情况（目标 1B）。 2）我们将研究HIV/HBV诱导的T细胞 损害。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 使用单细胞转录组学研究 HBV/HIV 共感染期间抗原特异性 CD8 T 细胞的损伤（目的 2A）。我们还将评估阻断 INFAR1 对 HIV/HBV 诱导的免疫损伤的影响。我们将监控 HIV 和 HBV 特异性 T 细胞的 T 细胞耗竭功能的逆转以及 IFN 诱导的功能分析 HBV/HIV 共感染期间抗原特异性 CD8 T 细胞受损（目标 2B）。 3）我们将调查如何 HIV-1 感染会增加人源化小鼠体内肝脏中 HBV 诱导的致病性巨噬细胞的数量（目标 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（目标 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 诱导的肝病 人源化小鼠（目标 3C）。 因此，我们将利用我们在 HIV-1/HBV (Su) 病毒学和免疫学方面广泛的互补专业知识， 肝炎病毒（Ploss）并在人源化小鼠技术中实现了令人兴奋的目标。我们的工作将会取得进展 通过证明一种新型小动物模型可以成功地用于 HBV 和 HIV-1 研究领域 了解 HIV/HBV 合并感染和免疫反应，并建立相关肝脏疾病的治疗模型。

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.