Novel humanized mouse model of mucosal immunity

新型人源化小鼠粘膜免疫模型

• 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
• 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

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.

项目摘要 尽管人源化小鼠已成功用于HIV感染、癌症和肿瘤的体内研究， 免疫疗法、人类初始免疫应答和适应性免疫应答仍不理想，交叉反应性有限 鼠和人细胞因子之间的差异被认为是一个关键的促成因素。因此，人体粘膜 免疫系统没有完全发育，缺少几个组成部分，包括人 肺上皮细胞在其自然肺环境中。为了解决这些研究差距，我们提出了一个综合的 基因和细胞编辑方法开发下一代人源化小鼠用于人类研究 粘膜免疫我们的方法有两个目标：在目标1中，我们将构建和认证hNSGF- 用于人免疫细胞发育的SGM 3-IL 6-TSLP-TSLPR小鼠。我们将产生表达一种 完整的人TSLP受体并检查与人造血祖细胞（HPC）的植入 通过测量骨髓、脾脏、肠道、呼吸道、肺和血液中免疫细胞的细胞组成。 为了检查人类免疫功能，我们将使用流感疫苗（Flumist）的粘膜制剂作为免疫调节剂。 免疫触发物和抗原来源，并分析血液中的细胞因子反应 和脾;人细胞迁移到粘膜部位、脾和骨髓;以及疫苗抗原的诱导- 脾脏中的特异性T和B细胞。目的二：构建人源化气道上皮细胞，并对其进行鉴定。 hNSGF-SGM 3-IL 6-TSLP-TSLPR小鼠。我们将研究人类气道上皮细胞的发育， 将人支气管上皮祖细胞移植到NSGF-SGM 3-IL 6-TSLP-TSLPR小鼠（人源化或 不使用来自同一供体的HPC）通过人特异性靶的组织免疫荧光染色， HLA I类。为了建立人气道上皮细胞的功能，我们将用活的 流感病毒并测量包括IL-33在内的物种特异性警报素的诱导; I型干扰素特征 作为其体内功能以及组织组成和吸引力的量度 人体免疫细胞与人体气道上皮细胞的结合。我们的假设是一个完整的TSLP信号通路 并且人上皮细胞将通过促进串扰来改善人源化小鼠中的人粘膜免疫 间质细胞和免疫细胞之间的联系。这个模型可以用于i。粘膜疫苗开发，以及 二.其他呼吸道病毒的研究，包括SARS-CoV-2，它需要人类特异性受体， 建立感染。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， 在遗传可变供体的背景下体内粘膜免疫以及使粘膜免疫的研究成为可能。 佐剂、过敏原、疫苗和生物制剂。