Uncovering Basic Signaling Mechanisms in NK Cells in Mice and Humans

描述(申请人提供)：自然杀伤(NK)细胞是先天免疫系统的淋巴细胞，通过裂解肿瘤和病原体感染的细胞来保护我们。最近的实验表明，在受到病毒感染(如CMV)挑战的小鼠和人类中，产生了长寿命的“记忆”NK细胞，类似于适应性免疫系统中的记忆淋巴细胞。我们试图开发一种具有预测能力的定量机制框架，用于预测小鼠和人类的自然杀伤细胞(包括记忆中的自然杀伤细胞)中的信号和激活，方法是将计算机模拟和湿实验室实验相结合。以统计物理、非线性动力学和多元统计为基础的计算机建模将与标准生化和多参数单细胞质量细胞仪(CyTOF质量细胞仪)实验相结合，这些实验针对工程化的人类NK细胞系(NKL)以及来自小鼠和人类的幼稚和“记忆”NK细胞。我们将研究三个相互重叠的目标：目标1：建立一个定量机制框架，用于分析激活和抑制CMV编码的配体刺激的NK细胞的信号和激活。目的2：确定小鼠幼稚NK细胞和“记忆”NK细胞信号机制的差异。目的3：确定小鼠和人类“记忆”NK细胞信号机制的差异。在这些目标的成功完成后，我们将拥有一个研究小鼠和人类NK细胞信号的定量预测机制框架。从拟议的项目中获得的系统水平的知识也可以用于理解其他免疫细胞中不同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.