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

不同免疫球蛋白结晶区(Fc区)相互作用的重要性 BRED建立中效应性免疫细胞表达的Fc-γ异构体及其排列 人们越来越认识到对流感病毒的免疫力。然而，它的翻译价值 在小鼠身上对这些途径的研究在一定程度上受到主要物种数量差异的限制， Fcγ受体的结构和表达模式，特别是低亲和力受体聚集在 1号染色体。类似地，尽管老鼠和人类的免疫球蛋白基因都编码四种免疫球蛋白 恒定区基因从而产生四种免疫球蛋白同工型，物种间存在差异 这使得很难将小鼠的同源基因分配给人类免疫球蛋白恒定区基因。这些物种 差异也限制了老鼠作为评估试剂的临床前工具的使用。 例如疫苗和人源化的单抗。为了解决这个问题，我们有 产生的小鼠编码小鼠免疫球蛋白受体的三个座位，FcɣRII/III/IV，FcɣR1a， 和FcRN(免疫球蛋白转运体)，通过共线替换人源化。人性化的小鼠 Fc-ɣ受体及其衍生系仅表达三个低亲和力FCGR激活中的一个 受体基因FCGR2A、FCGR2C或FCGR3A将被用来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，可以在免疫球蛋白同型抗体产生的动物身上研究受体。 病毒具有人类Fc区，确保Fc-FcɣR相互作用与在 人类。我们通过使~200kb的小鼠免疫球蛋白恒定区人性化来解决这一限制， 以及卡帕轻链的恒定区域。作为小鼠的胚胎干细胞 人源化的FCGR基因被用于这一基因组工程，产生的小鼠将 不仅产生人类免疫球蛋白亚型，而且这些亚型还会与人类效应器相互作用。 Fcɣ受体对效应细胞群体的影响。这些动物将为评估 人源单抗的有效性以及用于定义其参与的FC-FCɣR通路的有效性 调节病毒暴露后疾病的发病机制和/或提高机体免疫力 接种疫苗的动物。

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.