Generation of novel histoculture methods for studying human NK cell development

研究人类 NK 细胞发育的新型组织培养方法的产生

基本信息

项目摘要

Human natural killer (NK) cells play a critical role in the control of viral infection, particularly herpesviral infections such as Epstein-Barr virus (EBV). Their primary function of killing of virally infected target cells is mediated by the contact-dependent lysis through directed secretion of perforin and granzymes. Despite the demonstrated importance of their development and function, our understanding of how NK cells interact with other cells within tissue is limited by a lack of satisfying models for studying human immunity live and in situ. Significant challenges arise when trying to visualize cells within tissue, however it is highly relevant to human disease to find ways to better define how particularly innate immune cells migrate and exert their cytotoxic function. Our previous research has used 2D cell culture systems to define the importance of cell migration in human NK cell development. In this application, we propose the adoption of previously methods of secondary lymphoid tissue (tonsil) histoculture as a model to study human NK cell migration and function within an in vivolike tissue microenvironment using fast, low-toxicity imaging. We will combine our expertise in human developmental immunology with our technical skills in cutting-edge microscopy and image analysis to visualize human NK cells within autologous tissue and measure migration in 3 dimensions. We will additionally use multi-parametric imaging mass cytometry to localize cells within tissue (Aim 1). Further, we will extend our model to include infection of the tonsils with EBV, which will allow us to measure differences in cell migration and function between infected and uninfected tissue. By integrating infection of tissue histoculture with human EBV we will enable the study of human NK cells killing physiologically relevant targets within tissue (Aim 2). The generation of this novel model will advance future studies of human immunity and enable unprecedented insight into the behavior of immune cells within their tissue microenvironment.
人类自然杀伤(NK)细胞在控制病毒感染,特别是疱疹病毒感染中起着关键作用 感染,如EB病毒(EBV)。它们杀死病毒感染的靶细胞的主要功能是 介导的接触依赖性裂解通过直接分泌的穿孔素和颗粒酶。尽管 证明了它们的发展和功能的重要性,我们对NK细胞如何与 组织内的其它细胞由于缺乏用于活体和原位研究人类免疫的令人满意的模型而受到限制。 当试图可视化组织内的细胞时,出现了重大挑战,然而它与人类高度相关。 疾病,以找到更好地定义特别是先天免疫细胞如何迁移和发挥其细胞毒性的方法。 功能我们以前的研究已经使用2D细胞培养系统来定义细胞迁移在细胞增殖中的重要性。 人NK细胞发育。在本申请中,我们建议采用以前的方法, 淋巴组织(扁桃体)组织培养作为研究人NK细胞在体内样细胞内迁移和功能的模型 组织微环境使用快速,低毒性成像。我们将联合收割机结合我们在人类 发展免疫学与我们的技术技能,在尖端的显微镜和图像分析可视化 自体组织中的人NK细胞并测量三维迁移。我们还将使用 多参数成像质谱细胞术以定位组织内的细胞(Aim 1)。此外,我们将扩大我们的 模型包括EBV感染的扁桃体,这将使我们能够测量细胞迁移的差异 和功能之间的关系。通过组织培养与人感染相结合, 我们将能够研究人NK细胞杀死组织内生理学相关靶点(目的2)。 这种新模型的产生将推动未来对人类免疫的研究,并使前所未有的 深入了解免疫细胞在其组织微环境中的行为。

项目成果

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Emily Margaret Mace其他文献

Emily Margaret Mace的其他文献

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{{ truncateString('Emily Margaret Mace', 18)}}的其他基金

Defining the functional role of CD56 on human natural killer cells
定义 CD56 对人类自然杀伤细胞的功能作用
  • 批准号:
    10735537
  • 财政年份:
    2023
  • 资助金额:
    $ 8.1万
  • 项目类别:
DETERMINING THE ROLE OF THE REPLICATIVE HELICASE IN HUMAN NK CELL DEVELOPMENT
确定复制解旋酶在人类 NK 细胞发育中的作用
  • 批准号:
    10468048
  • 财政年份:
    2018
  • 资助金额:
    $ 8.1万
  • 项目类别:
DEFINING THE ROLE OF CELL MIGRATION IN HUMAN NK CELL DIFFERENTIATION
定义细胞迁移在人类 NK 细胞分化中的作用
  • 批准号:
    9897729
  • 财政年份:
    2018
  • 资助金额:
    $ 8.1万
  • 项目类别:
DEFINING THE ROLE OF CELL MIGRATION IN HUMAN NK CELL DIFFERENTIATION
定义细胞迁移在人类 NK 细胞分化中的作用
  • 批准号:
    10190802
  • 财政年份:
    2018
  • 资助金额:
    $ 8.1万
  • 项目类别:
DEFINING THE ROLE OF CELL MIGRATION IN HUMAN NK CELL DIFFERENTIATION
定义细胞迁移在人类 NK 细胞分化中的作用
  • 批准号:
    10410409
  • 财政年份:
    2018
  • 资助金额:
    $ 8.1万
  • 项目类别:
DETERMINING THE ROLE OF THE REPLICATIVE HELICASE IN HUMAN NK CELL DEVELOPMENT
确定复制解旋酶在人类 NK 细胞发育中的作用
  • 批准号:
    9790924
  • 财政年份:
    2018
  • 资助金额:
    $ 8.1万
  • 项目类别:

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