Behavioral dynamics of Langerhans cells in skin

皮肤朗格汉斯细胞的行为动力学

基本信息

批准号：
7474706
负责人：
AKIRA TAKASHIMA
金额：
$ 31.05万
依托单位：
UNIVERSITY OF TOLEDO HEALTH SCI CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-09-15 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7474706
关键词：
Amoeba genus Animals Antigens Behavior Behavioral Behavioral Mechanisms Biology Breeding Cell Adhesion Molecules Cell Maturation Cells Cellular biology Color Condition Data Set Dendrites Dendritic Cells Dermal Detection Dinitrofluorobenzene DsRed Ear Engineering Exhibits Exposure to Extracellular Space Four-dimensional Frequencies Functional disorder Habitats Image Immunobiology Immunologic Monitoring In Situ In Vitro Individual Infection Inflammatory Kinetics Knock-in Mouse Knockout Mice Knowledge Langerhans cell Laser Scanning Microscopy Lateral Life Location Measurement Measures Mediator of activation protein Medical Surveillance Microscopic Modeling Motion Movement Mus Organ Culture Techniques Play Population Positioning Attribute Process Proteins Purpose Rate Research Personnel Role Sampling Signal Transduction Skin Staining method Stains Standards of Weights and Measures Statistically Significant Stimulus Streptococcus pyogenes System T-Lymphocyte Testing Thinking Three-Dimensional Image Three-Dimensional Imaging Time Tissue Fixation Tissue Sample Topical application Transgenic Mice base cell behavior cell motility cell preparation chemokine clinically relevant cytokine in vivo insight irradiation keratinocyte langerin lymph nodes microbial migration multi-photon novel progenitor promoter red fluorescent protein research study response skin disorder two-photon ultraviolet uptake

项目摘要

DESCRIPTION (provided by applicant): Langerhans cells (LCs) are immature dendritic cell (DC) subsets that perform two key surveillance tasks (antigen sampling and detection of danger signals) at the environmental interface. Our objective is to define behavioral mechanisms by which LCs achieve these tasks in their natural habitat (i.e., skin). For this purpose, we have developed novel imaging systems to acquire 4D images of LCs in living mice. In standard models of LC maturation, LCs exhibited dSEARCH motion (characterized by extension and retraction of dendrites), amoeba-like lateral migration, vertical migration, and LC-LC contact formation. Our aims are: J_) To define steady-state LC behaviors. LCs will be visualized in the l-Ap-EGFP knock-in mice (in which EGFP fluorescent signals are expressed by all DC subsets) and in the langerin-EGFP knock-in mice (in which only LCs express EGFP). We will record 3D images of EGFP* LCs every 2 min for 4 h by "time-lapse" multi- photon laser scanning microscopy in anesthetized mice to measure spaciotemporal kinetics of individual motile activities of EGFP+ LCs. We will study steady-state LC turnover by acquiring images of EGFP+ LCs in the same microscopic fields at different time points ("intermittent" imaging). By comparing the cellular movement recorded in the l-Ap-EGFP knock-in versus langerin-EGFP knock-in mice, we will assess the behaviors of other DC subsets in the skin. 2) To identify LC behavioral responses to pathological stimuli. We will study the turnover and motile activities of EGFP+ LCs fluorescent protein (dsRed). The EGFP knock-in mice will be cross-bred with: a) IL-1p promoter-driven dsRed-transgenic mice to study LC maturation (red signals) and LC motility (green signals) independently, and b) K14 promoter-driven dsRed-transgenic mice to study dynamic interactions of LCs (green) with neighboring keratinocytes (red). Our study should provide important insights into the behavioral biology of LCs and the pathophysiology of skin disease in which LCs play pathogenic or protective roles.

描述（由申请人提供）：Langerhans细胞（LCS）是未成熟的树突状细胞（DC）子集，它们在环境界面执行了两个关键的监视任务（抗原采样和危险信号的检测）。我们的目标是定义LCS在其自然栖息地（即皮肤）中实现这些任务的行为机制。为此，我们开发了新颖的成像系统，以获取活小鼠中LC的4D图像。在LC成熟的标准模型中，LCS表现出DSEarch运动（以树突的扩展和缩回为特征），变形虫样的横向迁移，垂直迁移和LC-LC接触形成。我们的目标是：J_）定义稳态LC行为。 LC将在L-AP-EGFP敲入小鼠（所有DC子集表达EGFP荧光信号）和Langerin-EGFP敲入小鼠（其中只有LCS Expears EGFP）中可视化LC。我们将通过麻醉小鼠中的“延时”多子激光扫描显微镜每2分钟记录每2分钟的EGFP* LCS的3D图像，以测量EGFP+ LCS单个运动活性的时空动力学。我们将通过在不同时间点（“间歇性”成像）在相同的显微镜字段中获取EGFP+ LC的图像来研究稳态LC。通过比较L-AP-EGFP敲入与Langerin-EGFP敲入小鼠中记录的细胞运动，我们将评估皮肤中其他DC子集的行为。 2）确定对病理刺激的LC行为反应。我们将研究EGFP+ LCS荧光蛋白（DSRED）的营业额和运动活性。 EGFP敲击小鼠将与以下杂交：a）IL-1P启动子驱动的DSRED dSRED-转基因小鼠独立研究LC成熟（红色信号）和LC运动性（绿色信号），以及b）K14启动子驱动子驱动的dsred dsred dsred dsred-transgenic小鼠与LCS（绿色）（绿色）（绿色）相同的动态相互作用。我们的研究应为LCS的行为生物学以及LCS发挥致病性或保护作用的皮肤疾病的病理生物生物学提供重要见解。