Yao Yi, Matsushima Hironori, Ohtola Jennifer A, Geng Shuo, Lu Ran, Takashima Akira
Department of Medical Microbiology and Immunology, University of Toledo College of Medicine, Toledo, OH 43614.
Department of Medical Microbiology and Immunology, University of Toledo College of Medicine, Toledo, OH 43614
J Immunol. 2015 Feb 1;194(3):1211-24. doi: 10.4049/jimmunol.1402018. Epub 2014 Dec 19.
Rapid enhancement of phagocyte functionality is a hallmark of neutrophil priming. GeneChip analyses unveiled elevated CD54, dectin-2, and IL-1β mRNA expression by neutrophils isolated from inflammatory sites. In fact, CD54 and dectin-2 protein expression was detected on neutrophils recovered from skin, peritoneal, and lung inflammation lesions but not on those in bone marrow or peripheral blood. Neutrophils increased CD54 and dectin-2 mRNA during migration in Boyden chambers and acquired CD54 and dectin-2 surface expression after subsequent exposure to GM-CSF. Neutrophils purified from IL-1β promoter-driven DsRed-transgenic mice acquired DsRed signals during cell migration or exposure to GM-CSF. CD54 and dectin-2 were expressed by DsRed(+) (but not DsRed(-)) neutrophils in GM-CSF-supplemented cultures, and neutrophils recovered from inflammatory sites exhibited strong DsRed signals. The dynamic process of neutrophil priming was studied in chemically induced inflammatory skin lesions by monitoring DsRed expression using confocal microscopy. A majority (>80%) of Ly6G(+) neutrophils expressed DsRed, and those DsRed(+)/Ly6G(+) cells exhibited crawling motion with a higher velocity compared with their DsRed(-)/Ly6G(+) counterparts. This report unveils motile behaviors of primed neutrophils in living animals. We propose that neutrophil priming occurs in a sequential manner with rapid enhancement of phagocyte functionality, followed by CD54 and dectin-2 mRNA and protein expression, IL-1β promoter activation, and accelerated motility. Not only do these findings provide a new conceptual framework for our understanding of the process of neutrophil priming, they also unveil new insights into the pathophysiology of many inflammatory disorders that are characterized by neutrophil infiltration.
吞噬细胞功能的快速增强是中性粒细胞预激活的一个标志。基因芯片分析显示，从炎症部位分离出的中性粒细胞中，CD54、dectin-2和IL-1β mRNA表达升高。事实上，在从皮肤、腹膜和肺部炎症病变中回收的中性粒细胞上检测到了CD54和dectin-2蛋白表达，但在骨髓或外周血中的中性粒细胞上未检测到。中性粒细胞在博伊登小室迁移过程中CD54和dectin-2 mRNA增加，并在随后暴露于GM-CSF后获得CD54和dectin-2表面表达。从IL-1β启动子驱动的DsRed转基因小鼠中纯化的中性粒细胞在细胞迁移或暴露于GM-CSF期间获得DsRed信号。在添加GM-CSF的培养物中，DsRed(+)（而非DsRed(-)）中性粒细胞表达CD54和dectin-2，并且从炎症部位回收的中性粒细胞表现出强烈的DsRed信号。通过共聚焦显微镜监测DsRed表达，在化学诱导的炎症性皮肤病变中研究了中性粒细胞预激活的动态过程。大多数（>80%）Ly6G(+)中性粒细胞表达DsRed，与DsRed(-)/Ly6G(+)细胞相比，那些DsRed(+)/Ly6G(+)细胞表现出更高速度的爬行运动。本报告揭示了活体动物中预激活中性粒细胞的运动行为。我们提出，中性粒细胞预激活以吞噬细胞功能的快速增强、随后CD54和dectin-2 mRNA及蛋白表达、IL-1β启动子激活和运动加速的顺序方式发生。这些发现不仅为我们理解中性粒细胞预激活过程提供了一个新的概念框架，还揭示了许多以中性粒细胞浸润为特征的炎症性疾病病理生理学的新见解。
