Modulation of T Cell Function by Patterning of Costimulatory Ligands

描述(由申请人提供)：T细胞的激活是由T细胞和抗原提呈细胞之间的动态界面介导的，称为免疫突触。这些研究的长期目标是了解免疫突触内分子的组织如何驱动细胞功能，以及调节这一过程的分子机制。初步研究表明，T细胞受体和CD28共刺激配体的微米级分离增强了自然小鼠CD4+T细胞产生IL2的能力。建议的研究试图确定从受体参与到IL2分泌的细胞信号通路中的关键点，作为对细胞识别和响应共刺激信号分离的潜在机制的调查。拟议的研究将使用两种方法来定义含有TCR和CD28配体的多组分表面。首先，微接触打印将被用来定义TCR(CD3)和CD28的激活抗体的共局定位和分离模式。其次，膜微流控技术将被用来创建支持的脂质双层的排列区域，I-Ek和CD80(分别是TCR和CD28的天然配体)被拴在这些区域上。免疫学和活体成像方法将被用来识别由配体分离引起的信号和迁移行为的差异。随后的研究将重点放在所识别的蛋白质的特定生物物理呈现上，以努力确定信号分离所引发的分子机制。这些研究的成功完成将为细胞整合细胞外环境的多种线索的机制提供新的见解，这一能力将影响理解细胞如何组织成各种重要的生理系统。此外，在这里获得的知识可能在改进T细胞群体的体外扩增方面有应用，目前这是使用珠子和其他简单地包裹着激活抗体的结构进行的；理解这些信号的图案化和调节T细胞功能如何立即适用于改进这些系统。项目简介免疫系统的适当功能对各种疾病以及治疗潜力具有强大的影响。这项拟议的研究试图了解改变呈现给T细胞的生物分子的分布如何影响它们发育为完全激活的细胞，这是在许多T细胞相互作用中观察到的现象。成功完成这些研究将有助于更好地了解免疫系统的功能，并可以改进定制免疫反应的设备和方法。

DESCRIPTION (provided by applicant): T cell activation is mediated by a dynamic interface between T cells and antigen presenting cells, termed the immunological synapse. The long-term goal of these studies is to understand how the organization of molecules within the immunological synapse drives cell function as well as the molecular mechanisms that mediate such processes. Preliminary studies demonstrate that micrometer-scale segregation of T cell receptor and CD28 costimulatory ligands enhances IL2 production by na¿ve murine CD4+ T cells. The proposed studies seek to identify key points of modulation in the cell signaling pathways that lead from receptor engagement to IL2 secretion, as a survey for potential mechanisms by which cells recognize and respond to the segregation of costimulatory signals. The proposed studies will use two approaches for defining multicomponent surfaces containing ligands to TCR and CD28. First, microcontact printing will be used to define colocalized and segregated patterns of activating antibodies to TCR (CD3) and CD28. Second, membrane microfluidics will be used to create aligned regions of supported lipid bilayer to which I-Ek and CD80 (natural ligands to TCR and CD28, respectively) are tethered. Immunological and live imaging approaches will be used to identify differences in signaling and migratory behavior that are induced by segregation of ligands. Subsequent studies will focus on the specific biophysical presentation of proteins that are identified, in an effort to identify the molecular mechanisms that are being invoked by signal segregation. Successful completion of these studies will provide new insight into mechanisms by which cells integrate multiple cues of the extracellular environment, an ability that impacts on understanding how cells organize into a variety of physiologically important systems. Moreover, the knowledge gained here may have application in improving the ex vivo expansion of T cell populations, which is currently carried out using beads and other structures that are simply coated with activating antibodies; understanding how patterning of these signals and modulate T cell function is immediately applicable to improving these systems.7. Project Narrative Proper function of the immune system has strong impacts on a variety of diseases as well as therapeutic potential. The proposed study seeks to understand how changing the distribution of biomolecules presented to T cells influences their development into fully activated cells, a phenomenon observed in many T cell interactions. Successful completion of these studies will lead to a better understanding of immune system function and can lead to improved devices and methods for tailoring immune responses.