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 结晶 (Fc) 区域片段之间相互作用的重要性 效应免疫细胞表达的亚型和 FcγR 阵列在建立广泛的 人们越来越认识到对流感病毒的免疫力。然而，翻译价值 对小鼠这些途径的研究部分受到数量上主要物种差异的限制， FcγR 的结构和表达模式，特别是聚集在其上的低亲和力受体 1号染色体。类似地，虽然小鼠和人类的IgG基因座都编码四种IgG 恒定区基因，从而产生四种 IgG 同种型，物种之间存在差异 使得很难将小鼠直系同源物分配给人类 IgG 恒定区基因。这些物种 差异也限制了小鼠作为评估试剂的临床前工具的使用 例如疫苗和人源化单克隆抗体（mAb）。为了解决这个问题，我们有 生成的小鼠中编码小鼠 IgG 受体的三个基因座 FcɣRII/III/IV、FcɣR1a、 和 FcRn（IgG 转运蛋白）通过同线性替换进行人源化。小鼠拟人化 FCɣR 和仅表达三种低亲和力 FCGR 激活之一的衍生系 受体基因 FCGR2A、FCGR2C 或 FCGR3A 将用于评估这些基因的作用 抗体介导的流感保护中的受体。人类Fc的贡献 理想情况下，应该在动物中研究受体，其中 IgG 同种型响应于 病毒具有人类 Fc 区，确保 Fc-FCɣR 相互作用模仿在 人类。我们通过对约 200 kb 的小鼠 IgH 恒定区进行人源化来解决这一限制， 以及 kappa 轻链的恒定区。作为来自小鼠的胚胎干细胞 FCGR基因人源化用于该基因组工程，产生的小鼠将 不仅产生人类 IgG 同工型，而且这些同工型还会与人类效应子相互作用 FCɣRs 对效应细胞群的影响。这些动物将为评估 人类单克隆抗体的有效性以及定义 Fc-FcɣR 通路的作用 调节病毒暴露后疾病的发病机制和/或提高免疫力 已接种疫苗的动物。