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

项目摘要 与乙肝病毒和HIV-1混合感染很常见，HIV混合感染会加剧病毒性肝炎的进展 并加速肝病的发展。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 用乙肝病毒和HIV-1复制类似人类的感染。这两种病毒的宿主范围都很窄，感染是 主要局限于人类和其他类人猿。我们的实验室(普洛斯和苏)已经开发出新的人源化小鼠 与人类肝脏和人类免疫细胞(FNRGF/A2-Hu Hep/HSC小鼠)共移植，与人类 肝细胞和改良的人髓系细胞/DC和NK细胞亚群在常规治疗中表达不足 人性化的老鼠模型。我们已经证明，这些改进可以产生疫苗诱导的免疫原性。 与接种了同样明确的YFV-17D疫苗的人相似。我们 将利用这些有希望的突破来调查以下内容：1)我们将感染FNRGF/A2的队列- HU小鼠植入人类白细胞抗原相合的肝细胞和携带乙肝病毒和/或病毒的人造血细胞 HIV-1。我们将纵向监测和量化乙肝病毒和艾滋病毒病毒血症以及免疫激活，包括抗原- 特异性CD8 T细胞与肝病进展(目标1A)。我们还将对乙肝病毒/艾滋病毒的队列进行分析。 接受HAART(包括乙肝病毒聚合酶抑制剂)治疗的混合感染小鼠，以抑制HIV-1 和乙肝病毒，以准确地模拟临床相关情况(目标1B)。2)我们将研究艾滋病毒/乙肝病毒诱导的T细胞 减损。我们将监测HIV和HBV特异性T细胞上的T细胞耗竭标志物，并分析其功能 应用单细胞转录组学技术研究乙肝病毒/艾滋病病毒混合感染过程中抗原特异性CD8 T细胞的损伤 2a)。我们还将评估阻断INFAR1对艾滋病毒/乙肝病毒诱导的免疫损伤的影响。我们会监控 人类免疫缺陷病毒和乙型肝炎病毒特异性T细胞的T细胞衰竭功能逆转及干扰素诱导的功能变化 乙肝病毒/艾滋病毒混合感染期间抗原特异性CD8 T细胞的损害(目标2B)。3)我们将调查如何 HIV-1感染增加体内人源化小鼠肝脏中由乙肝病毒诱导的致病巨噬细胞(目标3A)。 我们还将研究艾滋病毒和乙肝病毒如何与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艾滋病毒/乙肝病毒诱导的肝病。 人源化小鼠(AIM 3C)。 因此，我们将充分利用我们在HIV-1/HBV(SU)病毒学和免疫学方面的广泛免费专业知识， 肝炎病毒(普洛斯)和人性化小鼠技术达到了激动人心的目的。我们的工作将会向前推进 通过证明一种新的小动物模型可以成功地用于 了解艾滋病毒/乙肝病毒混合感染和免疫反应，以及相关肝病的模型治疗。

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.