Uncovering Basic Signaling Mechanisms in NK Cells in Mice and Humans

描述（由申请人提供）：自然杀伤（NK）细胞是先天免疫系统的淋巴细胞，通过溶解肿瘤和病原体感染的细胞来保护我们。最近的实验表明，在病毒感染（如巨细胞病毒）的挑战下，小鼠和人类产生了长寿的“记忆”NK细胞，类似于适应性免疫系统中的记忆淋巴细胞。我们试图开发一个定量机制框架，通过使用硅模型和湿实验室实验的协同结合，预测NK细胞（包括小鼠和人类的“记忆”NK细胞）的信号传导和激活能力。基于统计物理、非线性动力学和多元统计学的计算机建模将与标准生化和多参数单细胞细胞计数（CyTOF）实验相结合，实验对象是工程人类NK细胞系（NKLs），以及来自小鼠和人类的原始NK细胞和“记忆”NK细胞。我们将研究三个重叠的目标：目标1：建立一个定量的机制框架来分析激活和抑制cmv编码配体刺激的NK细胞的信号传导和激活。目的2：确定小鼠初始NK细胞和“记忆”NK细胞之间信号传导机制的差异。目的3：确定小鼠和人类“记忆”NK细胞之间信号传导机制的差异。在成功完成这些目标后，我们将有一个定量预测机制框架来研究小鼠和人类NK细胞信号传导。系统水平的知识也可以应用于理解其他免疫细胞中由异质受体配体相互作用调节的激活机制。

DESCRIPTION (provided by applicant): Natural Killer (NK) cells are lymphocytes of the innate immune system that defend us by lysing tumor and pathogen-infected cells. Recent experiments showed generation of long-lived "memory" NK cells in mouse and humans challenged by viral infections (such as CMV), similar to that of memory lymphocytes in the adaptive immune system. We seek to develop a quantitative mechanistic framework with predictive powers for signaling and activation in NK cells including "memory" NK cells in mice and humans by using a synergistic combination of in silico modeling and wet lab experiments. The in silico modeling rooted in statistical physics, non-linear dynamics, and multivariate statistics will be combined with standard biochemical and multi-parametric single cell mass cytometry (CyTOF mass cytometer) experiments on engineered human NK cell lines (NKLs), and naive and "memory" NK cells from mice and humans. We will investigate three overlapping aims: Aim 1: Develop a quantitative mechanistic framework for analyzing signaling and activation of NK cells stimulated by activating and inhibitory CMV-encoded ligands. Aim 2: Determine differences in signaling mechanisms between naive and "memory" NK cells in the mouse. Aim 3: Determine differences in signaling mechanisms between mouse and human "memory" NK cells. At the successful completion of the aims we will have a quantitative predictive mechanistic framework for studying NK cell signaling in mice and humans. The systems level knowledge gained from the proposed projects can also be applied in understanding mechanisms of activation regulated by heterogeneous receptor ligand interactions in other immune cells.