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 尽管人性化小鼠已成功用于HIV感染，癌症和 免疫疗法，幼稚和适应性免疫回报保持不佳，交叉反应有限 在鼠和人类细胞因子之间被认为是关键因素。结果，人类粘膜 免疫系统尚未完全发达，并且缺少几个组成部分，包括人类的存在 肺上皮细胞在其自然肺部环境中。为了解决这些研究差距，我们提出了一个合并的 遗传和细胞编辑方法开发了下一代人类小鼠的研究 粘膜免疫。我们的方法是两个目标结构的：在AIM 1中，我们将构建和凭证HNSGF- SGM3-IL6-TSLP-TSLPR小鼠用于人类免疫细胞发育。我们将产生表达的小鼠 人类造血祖细胞（HPC）完整的人类TSLP受体和检查工程师 通过测量骨髓，sleen，肠道，气道，肺和血液中免疫细胞的细胞组成。 为了检查人类免疫学功能，我们将使用粘膜疫苗（Flumist）的粘膜式作为一种 与过去一样，免疫触发和抗原来源和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；并诱导疫苗抗原 脾脏中的特异性T和B细胞。在AIM 2中，我们将在 HNSGF-SGM3-IL6-TSLP-TSLPR小鼠。我们将检查人类气道上皮的发展 NSGF-SGM3-IL6-TSLP-TSLPR小鼠中人支气管上皮祖细胞的移植 没有来自同一供体的HPC）通过组织免疫荧光染色的人类特异性靶标 HLA I级I。为了建立人类气道上皮细胞的功能，我们将挑战小鼠。 流感病毒并测量包括IL-33在内的特定规范警报的诱导；类型I干扰素签名 在人体上皮细胞中，作为其体内功能的量度，以及组织组成和吸引力 人类免疫细胞对人气道上皮的细胞。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性化小鼠的人类粘膜免疫 在基质细胞和免疫细胞之间。然后可以将此模型用于i。粘膜疫苗开发， ii。对包括SARS-COV-2在内的其他呼吸道病毒的研究，该病毒需要人类特异性受体 建立感染。一旦获得证书，我们的模型将为未来的机械研究奠定基础 在一般可变的供体的背景下，体内粘膜免疫，并能够研究粘膜 佐剂，过敏原，疫苗和生物制剂。