Novel humanized mouse model of mucosal immunity

新型人源化小鼠粘膜免疫模型

基本信息

批准号：
10591854
负责人：
Anna Karolina Palucka
金额：
$ 29.16万
依托单位：
JACKSON LABORATORY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-01-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10591854
关键词：
2019-nCoV Address Adjuvant Allergens Antibodies Antigens B-Cell Activation B-Lymphocytes Biological Blood Bone Marrow Bronchi CD34 gene Cell Communication Cell physiology Cells Clinic Clustered Regularly Interspaced Short Palindromic Repeats Colon Credentialing Dendritic Cells Development Disease Engraftment Environment Epithelial Cells Epithelium Extrinsic asthma FLT3 gene Fibroblasts FluMist Formulation Future Generations Genetic Genetic Engineering HIV Infections Helper-Inducer T-Lymphocyte Hematopoietic Hematopoietic stem cells Homeostasis Human Hypersensitivity IL6 gene IL7R gene Immune Immune response Immune system Immunodeficient Mouse Immunofluorescence Immunologic Immunotherapy In Vitro Infection Inflammation Inflammatory Innate Immune Response Interferon Type I Interleukin-6 Interleukins Knock-in Knock-in Mouse Knockout Mice Liver Lung Lymphoid Lymphoid Tissue Macrophage Macrophage Colony-Stimulating Factor Malignant Neoplasms Measures Modeling Mucosal Immune System Mucosal Immunity Mucous Membrane Mus Myeloid Cells National Institute of Allergy and Infectious Disease Natural regeneration Peripheral Blood Mononuclear Cell Phenotype Plasma Cells Play Population Pre-Clinical Model Property Proteins Research Respiratory Tract Infections Signal Pathway Signal Transduction Smooth Muscle Myocytes Source Spleen Stains Stem Cell Factor Stromal Cells Structure System Systemic infection T cell response T-Lymphocyte TSLP gene Testing Thymic Tissue Tissues Transgenic Organisms Transplantation United States National Institutes of Health Vaccine Antigen Vaccines Virus adaptive immune response adaptive immunity airway epithelium antigen-specific T cells bronchial epithelium cell motility cell regeneration cell type cross reactivity cytokine enzyme linked immunospot assay epithelial stem cell fetal genetic approach human stem cells humanized mouse immune function improved in vivo influenza infection influenza virus vaccine influenzavirus lung injury monocyte mouse development mouse model mucosal site mucosal vaccine next generation novel overexpression permissiveness receptor respiratory colonization respiratory smooth muscle respiratory virus response tissue injury translational immunology vaccine development

项目摘要

PROJECT SUMMARY Although humanized mice have been successfully used for in vivo studies of HIV infection, cancer and immunotherapies, human naïve and adaptive immune responses remain suboptimal, with limited cross-reactivity between murine and human cytokines considered as a key contributing factor. As a result, the human mucosal immune system is not fully developed, and several components are missing, including the presence of human lung epithelial cells in their natural lung environment. To address these research gaps, we propose a combined genetic and cellular editing approach to develop the next generation of humanized mice for studies of human mucosal immunity. Our approach is structured in two aims: in Aim 1, we will construct and credential hNSGF- SGM3-IL6-TSLP-TSLPR mice for human immune cell development. We will generate mice expressing a complete human TSLP receptor and examine engraftment with human hematopoietic progenitor cells (HPCs) by measuring cellular composition of immune cells in the bone marrow, spleen, gut, airways, lungs, and blood. To examine human immune function, we will use a mucosal formulation of influenza vaccine (Flumist) as an immune trigger and antigen source as we have done in the past and analyze cytokine responses in the blood and spleen; human cell migration to mucosal sites, spleen, and bone marrow; and induction of vaccine antigen- specific T and B cells in the spleen. In Aim 2, we will construct and credential humanized airway epithelium in hNSGF-SGM3-IL6-TSLP-TSLPR mice. We will examine the development of human airway epithelium upon transplant of human bronchial epithelial progenitor cells in NSGF-SGM3-IL6-TSLP-TSLPR mice (humanized or not with HPCs from the same donor) by tissue immunofluorescence staining of human-specific targets including HLA class I. To establish the functionality of human airway epithelial cells, we will challenge mice with live influenza virus and measure the induction of species-specific alarmins including IL-33; type I interferon signature in human epithelial cells as a measure of their functionality in vivo, as well as tissue composition and attraction of human immune cells to human airway epithelium. Our hypothesis is that a complete TSLP signaling pathway and human epithelial cells will improve human mucosal immunity in humanized mice by facilitating the crosstalk between stromal cells and immune cells. This model can then be used for i. mucosal vaccine development, and ii. studies of other respiratory viruses including SARS-CoV-2, which requires human-specific receptors to establish infection. Thus, once credentialed, our model will lay the groundwork for future mechanistic studies of mucosal immunity in vivo in the context of genetically variable donors as well as enable studies of mucosal adjuvants, allergens, vaccines, and biologicals.

项目概要尽管人源化小鼠已成功用于 HIV 感染、癌症和免疫疗法、人类初始免疫反应和适应性免疫反应仍然不理想，交叉反应性有限小鼠和人类细胞因子之间的差异被认为是一个关键因素。免疫系统尚未完全发育，缺少一些组件，包括人类免疫系统的存在为了解决这些研究空白，我们提出了一种组合方法。遗传和细胞编辑方法开发用于人类研究的下一代人源化小鼠我们的方法有两个目标：在目标 1 中，我们将构建并证明 hNSGF- 用于人类免疫细胞发育的 SGM3-IL6-TSLP-TSLPR 小鼠我们将产生表达 a 的小鼠。完整的人类 TSLP 受体并检查人类造血祖细胞 (HPC) 的植入通过测量骨髓、脾脏、肠道、气道、肺和血液中免疫细胞的细胞组成。为了检查人体的免疫功能，我们将使用流感疫苗（Flumist）的粘膜制剂作为正如我们过去所做的那样，免疫触发因素和抗原来源并分析血液中的细胞因子反应和脾脏；人类细胞迁移至粘膜部位、脾脏和骨髓以及疫苗抗原的诱导；在目标 2 中，我们将构建并验证脾脏中的人源化气道上皮细胞。 hNSGF-SGM3-IL6-TSLP-TSLPR 小鼠我们将检查人类气道上皮的发育。将人支气管上皮祖细胞移植到 NSGF-SGM3-IL6-TSLP-TSLPR 小鼠（人源化或不是来自同一供体的 HPC）通过人类特异性靶标的组织免疫荧光染色，包括 HLA I 类。为了确定人类气道上皮细胞的功能，我们将用活体细胞挑战小鼠流感病毒并测量物种特异性警报素（包括 IL-33 干扰素特征）的诱导；在人类上皮细胞中作为其体内功能以及组织组成和吸引力的衡量标准我们的假设是完整的 TSLP 信号通路。人类上皮细胞将通过促进串扰来改善人源化小鼠的人类粘膜免疫该模型可用于粘膜疫苗的开发，以及 ii. 对包括 SARS-CoV-2 在内的其他呼吸道病毒的研究，需要人类特异性受体来因此，一旦获得认证，我们的模型将为未来的机制研究奠定基础。在遗传变异供体的背景下进行体内粘膜免疫，并能够研究粘膜佐剂、过敏原、疫苗和生物制品。