Ly6 family members in neutrophil biology

项目摘要 中性粒细胞保护免受感染，但也介导炎性组织损伤。因此，在本发明中， 治疗上靶向嗜中性粒细胞将需要对其基本生物学的详细理解， 集中于中性粒细胞防御和致病功能可能发生分歧的领域。 在本研究的第一个周期中，我们探讨了两个相关的GPI连接的中性粒细胞表面 功能不佳的蛋白质，小鼠中的Ly6G和人类中的CD177。我们发现 这两种蛋白质在分子水平上与中性粒细胞表面β2整联蛋白顺式结合， 连接从而减弱嗜中性粒细胞迁移。利用实验潜力， 在小鼠炎症模型中，我们发现Ly6G连接选择性地减少了整合素介导的 中性粒细胞渗出向无菌触发物的典型迁移，使整合素非依赖性 对传染性刺激的迁移基本上不受干扰。我们的初步数据显示 Ly6G区分鼠嗜中性粒细胞内的亚表型，而在人类中， CD177阴性中性粒细胞在基因表达和潜在的细胞因子产生方面不同。一起 随着第一轮奖励的生产力，这些发现支持更深入的调查， 这些Ly6家族分子在中性粒细胞生物学中的作用。 我们提出了两个新的、独立的具体目标。目标一：寻求一种机制， Ly6G和CD 177改变中性粒细胞β2整合素功能，包括寻找新的内源性 反配体目的II开发了我们区分中性粒细胞亚型的初步RNAseq数据 基于Ly6G和CD177的表达，均来自健康供体以及成人和儿童 患有炎症性关节炎和短暂但强烈的炎症性血管炎川崎。 总之，这些研究将通过定义Ly6家族如何与中性粒细胞结合来促进对中性粒细胞的理解。 成员调节β 2整合素以控制细胞迁移和鉴定嗜中性粒细胞表型 反映在Ly6G和CD177的差异表达中。

Project Summary Neutrophils protect against infection but also mediate inflammatory tissue injury. As a result, targeting neutrophils therapeutically will require a detailed understanding of their basic biology, focused on domains wherein the defensive and pathogenic functions of neutrophils may diverge. In the first cycle of the present award, we explored two related GPI-linked neutrophil surface proteins of poorly-characterized function, Ly6G in mice and CD177 in humans. We showed that both proteins associate at a molecular level in cis with neutrophil surface β2 integrins and that their ligation thereby attenuates neutrophil migration. Taking advantage of the experimental potential of murine inflammatory models, we found that Ly6G ligation selectively reduced integrin-mediated migration typical for neutrophil diapedesis toward sterile triggers, leaving integrin-independent migration to infectious stimuli largely unperturbed. Our preliminary data now show evidence that Ly6G differentiates subphenotypes within murine neutrophils, while in humans CD177pos and CD177neg neutrophils differ in gene expression and potentially in cytokine production. Together with the productivity of the first cycle of the award, these findings support deeper investigation of the role of these Ly6-family molecules in neutrophil biology. We propose two new and independent specific aims. Aim I pursues the mechanisms by which Ly6G and CD177 alter neutrophil β2 integrin function, including a search for novel endogenous counterligands. Aim II develops our preliminary RNAseq data distinguishing neutrophil subtypes based on expression of Ly6G and CD177, both from healthy donors and from adults and children with inflammatory arthritis and the transient but intensely inflammatory vasculitis Kawasaki disease. Together, these studies will advance the understanding of neutrophils by defining how Ly6 family members regulate 2 integrins to control cell migration and identifying neutrophil phenotypes reflected in differential expression of Ly6G and CD177.