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 两者在分子水平上与嗜中性粒细胞表面β2整联蛋白顺式结合， 从而减弱嗜中性粒细胞迁移。利用小鼠的实验潜力 在炎症模型中，我们发现Ly6G连接选择性地减少了整合素介导的迁移， 典型的中性粒细胞向无菌触发物渗出，使整合素非依赖性迁移， 感染性刺激基本上不受干扰。我们的初步数据显示，Ly6G 在小鼠中性粒细胞中，CD177阳性和CD177阴性中性粒细胞中， 在基因表达和潜在的细胞因子产生方面存在差异。加上生产力的 在该奖项的第一个周期，这些发现支持这些Ly6家族的作用更深入的调查 中性粒细胞生物学中的分子 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Aim II开发了初步的RNAseq数据， 对Ly6G和CD177表达的影响，来自健康供体以及患有 炎性关节炎和短暂但强烈炎性血管炎川崎。 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，并通过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 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 that Ly6G differentiates subphenotypes within murine neutrophils, while in humans CD177pos and CD177neg neutrophils differ in gene expression and potentially 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 preliminary RNAseq data distinguishing neutrophil subtypes based on expression of Ly6G and CD177, from healthy donors as well as 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 by identifying neutrophil phenotypes reflected in differential expression of Ly6G and CD177.