Genetically humanized mice for modeling human Fc-receptor interaction during influenza infection

用于模拟流感感染期间人类 Fc 受体相互作用的基因人源化小鼠

• 批准号: 10117188
• 负责人: Beverly H Koller
• 金额: $ 19.44万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-03 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10117188
• 关键词: Address; Affinity; Alleles; Animals; Antibodies; Antibody-mediated protection; CRISPR/Cas technology; Cell Lineage; Cells; Chromosome 1; Crystallization; DNA; Development; Disease; ES Cell Line; Effector Cell; Ensure; Epitopes; Evaluation; FCGR2C gene; FCGR3A gene; Fc Receptor; Future; Generations; Genes; Genome engineering; Human; IgG Receptors; IgK; Immune; Immune response; Immunity; Immunoglobulin Constant Region; Immunoglobulin Fragments; Immunoglobulin G; Immunoglobulins; Individual; Influenza; Influenza A virus; Light; Mediating; Modeling; Modification; Monoclonal Antibodies; Mus; Mutation; Myelogenous; Natural Killer Cells; Orthologous Gene; Pathogenesis; Pathway interactions; Pattern; Phagocytosis; Population; Protein Isoforms; Reagent; Receptor Gene; Role; Structure; Transgenes; Transgenic Organisms; Vaccinated; Vaccines; Validation; Viral; Virus; anti-influenza; antibody-dependent cell cytotoxicity; blastocyst; chimeric antibody; constant region gene; effectiveness evaluation; embryonic stem cell; homologous recombination; human DNA; human model; human monoclonal antibodies; humanized monoclonal antibodies; humanized mouse; improved; influenza infection; influenzavirus; macrophage; monocyte; mutant; neonatal Fc receptor; pre-clinical; receptor; response; species difference; tool

The importance of interactions between the fragment of crystallization (Fc) region of various IgG isoforms and the array of FcγRs expressed by effector immune cells in establishment of broad immunity to influenza viruses is increasingly recognized. However, the translational value of studies of these pathways in mice is limited in part by major species differences in the number, structure and expression pattern of the FcγRs, particularly the low affinity receptors clustered on chromosome 1. Similarly, although both the mouse and the human IgG locus encode four IgG constant region genes and thus produce four IgG isotypes, divergence between the species has made it difficult to assign mouse orthologs to human IgG constant region genes. These species differences have also limited the use of the mouse as a preclinical tool for evaluating reagents such as vaccines and humanized monoclonal antibodies (mAbs).To address this, we have generated mice in which the three loci encoding mouse IgG receptors, FcɣRII/III/IV, FcɣR1a, and FcRn (the IgG transporter), are humanized by syntenic replacement. Mice humanized for the FCɣRs and derived lines expressing only one of the three low affinity FCGR activating receptor genes, FCGR2A, FCGR2C or FCGR3A, will be used to evaluate the role of these receptors in antibody-mediated protection against influenza. The contribution of human Fc receptors would ideally be studied in animals in which the IgG isotypes produced in response to virus have human Fc regions, ensuring that the Fc-FCɣR interactions mimic those observed in humans. We address this limitation by humanization of the ~200 kb mouse IgH constant region, as well as the constant region for the kappa light chains. As embryonic stem cells from mice humanized for the FCGR genes are used for this genome engineering, the mice generated will not only produce human IgG isoforms, but these isoforms will interact with human effector FCɣRs on effector cell populations.These animals will provide a model for evaluation of the effectiveness of human mAbs as well as for defining Fc-FcɣR pathways whose engagement modulates the pathogenesis of disease after viral exposure and/or improves immunity in vaccinated animals.

各种IgG的结晶片段（FC）区域之间相互作用的重要性 效应免疫细胞表达的同工型和FcγR的阵列在建立宽 越来越多地认识到对影响力病毒的免疫力。但是，翻译价值 在小鼠中对这些途径的研究部分受到该数量的主要物种差异的限制， FcγR的结构和表达模式，尤其是群集的低亲和力受体 类似地，染色体1。尽管小鼠和人IgG基因座都编码四个IgG 恒定区域基因，因此产生四个IgG同种型，该物种之间的差异具有 使小鼠直系同源物为人IgG恒定区域基因分配很难。这些物种 差异也限制了鼠标用作评估试剂的临床前工具的使用 例如疫苗和人源化的单克隆抗体（mabs）。要解决这个问题，我们有 生成的小鼠，其中三个基因座编码小鼠IgG受体，fcɣrii/iii/iv，fcɣr1a， 和FCRN（IgG转运蛋白）通过同义替换进行人性化。小鼠人性化 FCɣRS和派生线仅表达三个低亲和力FCGR激活之一 受体基因FCGR2A，FCGR2C或FCGR3A将用于评估这些作用 抗体介导的对影响力的受体。人类足球俱乐部的贡献 理想情况下，接收器将是在动物中研究的，其中IgG同种型响应于 病毒具有人类FC区域，确保FC-FCɣR相互作用模仿了 人类。我们通过人性化约200 kb小鼠ig恒定区域来解决这一限制， 以及Kappa轻链的恒定区域。作为小鼠的胚胎干细胞 用于FCGR基因的人源化用于该基因组工程，生成的小鼠将 不仅产生了人类IgG同工型，而且这些同工型将与人类效应子相互作用 这些动物将提供评估的模型 人mab的有效性以及定义互动的FC-FCɣR途径 调节病毒暴露后疾病的发病机理和/或改善免疫力 接种动物。