Helen L. and Martin S. Kimmel Center for Biology and Medicine in the Skirball Institute for Biomolecular Medicine and Department of Pathology, NYU School of Medicine, New York, NY 10016, USA.
Curr Top Microbiol Immunol. 2010;340:1-24. doi: 10.1007/978-3-642-03858-7_1.
Innate and adaptive immunity is dependent upon reliable cell-cell communication mediated by direct interactions of cell surface receptors with ligands integrated into the surface of apposing cells or bound directly to the surface as in complement deposition or antibody mediated recognition through Fc receptors. Supported lipid bilayers formed on glass surfaces offer a useful model system in which to explore some basic features of molecular interactions in immunological relevant contacts, which include signal integration and effector functions through immunological synapses and kinapses. We have exploited that lateral mobility of molecules in the supported planar bilayers and fluorescence microscopy to develop a system for measurement of two-dimensional affinities and kinetic rates in the contact area, which is of immunological interest. Affinity measurements are based on a modified Scatchard analysis. Measurements of kinetic rates are based on fluorescence photo bleaching after recovery at the level of the entire contact area. This has been coupled to a reaction-diffusion equation that allows calculation of on- and off-rates. We have found that mixtures of ligands in supported planar bilayers can effectively activate T lymphocytes and simultaneously allow monitoring of the immunological synapse. Recent studies in planar bilayers have provided additional insights into organization principles of cell-cell interfaces. Perennial problems in understanding cell-cell communication are yielding quantitative measurements based on planar bilayers in areas of ligand-driven receptor clustering and the role of the actin cytoskeleton in immune cell activation. A major goal for the field is determining quantitative rules involved in signaling complex formation by innate and adaptive receptor systems.
先天免疫和适应性免疫依赖于可靠的细胞间通讯，这种通讯是通过细胞表面受体与整合到相邻细胞表面的配体或直接与表面结合的配体（如补体沉积或通过 Fc 受体介导的抗体识别）的直接相互作用来介导的。在玻璃表面形成的支持脂双层提供了一个有用的模型系统，可用于探索免疫相关接触中分子相互作用的一些基本特征，包括通过免疫突触和 Kinapse 进行信号整合和效应功能。我们利用支持的平面双层中的分子的横向流动性和荧光显微镜来开发一种用于测量接触区域中二维亲和力和动力学速率的系统，该系统具有免疫学意义。亲和力测量基于改进的 Scatchard 分析。动力学速率的测量基于整个接触区域荧光漂白后的恢复。这与反应扩散方程耦合，该方程允许计算开和关的速率。我们发现，支持平面双层中的配体混合物可以有效地激活 T 淋巴细胞，同时允许监测免疫突触。最近在平面双层中的研究为细胞-细胞界面的组织原则提供了更多的见解。理解细胞间通讯的长期问题正在基于平面双层提供定量测量，这些测量涉及配体驱动的受体聚类以及肌动蛋白细胞骨架在免疫细胞激活中的作用。该领域的一个主要目标是确定先天和适应性受体系统形成信号复合物所涉及的定量规则。
