Mast cell activation on patterned lipid bilayers of subcellular dimensions
Mast cell activation on patterned lipid bilayers of subcellular dimensions
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DOI：
10.1021/la026314c
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发表时间：
2003-03-04
期刊：
影响因子：
3.9
通讯作者：
Baird, BA
中科院分区：
文献类型：
--
作者：
Orth, RN;Wu, M;Baird, BA
免疫系统中的T细胞、B细胞和其他特化细胞通过其质膜表面上抗原受体的聚集而被激活;表达细胞表面免疫球蛋白E（IgE）受体并在过敏反应中起作用的大鼠嗜碱性白血病（RBL）肥大细胞用作原型模型。为了研究细胞应答的信号传导途径，正在开发一种通用系统，用于在用基于聚合物的湿剥离方法制备的脂质双层的微米级图案上呈递抗原。在聚合物的光刻图案化之后并且在聚合物在溶液中被机械剥离之前，在硅基底上形成支撑的脂质双层。这形成了明确定义的双层模式，其可以包含半抗原化和荧光脂质，并且可以用于模拟细胞受体接合和响应的生物基质。对于该模型系统，将1，2-二棕榈酰-sn-甘油基-3-磷酸乙醇胺-N- [6- [（2，4-二硝基苯基）氨基]己酰基]（铵盐）（DNP-cap-DPPE）用于抗DNP IgE的特异性结合。如果肥大细胞表面受体（Fc是RI的一种成分）比细胞小得多，则IgE与肥大细胞表面受体结合，聚集并聚集在图案特征上。对于细胞大小或更大的图案化特征，细胞粘附到硅表面，受体朝向图案化脂质的边缘极化。具有特异性接合的IgE受体的RBL细胞被激活以经历可以用荧光或扫描电子显微镜分析的显著形态学变化。这种用于图案化抗原功能化脂质的新方法为抗原刺激提供了低至微米分辨率的空间控制，同时保留了质膜基底的特征，包括动力学和可变组成。这种方法提供了一种替代微接触印刷与细胞免疫学研究的一些增强的能力。
T Cells, B cells, and other specialized cells in the immune system are activated by clustering of antigenic receptors on their plasma membrane surfaces; rat basophilic leukemia (RBL) mast cells which express cell surface immunoglobulin E (IgE) receptors and operate in the allergic response serve as a prototypic model. To investigate signaling pathways of the cellular response, a versatile system is being developed for presenting antigens on micron scale patterns of lipid bilayers that are prepared with a polymer-based wet lift-off method. A supported lipid bilayer is formed on a silicon substrate after photolithographic patterning of a polymer and before the polymer is mechanically peeled away in solution. This forms well-defined bilayer patterns that can contain haptenated and fluorescent lipids and can be used to simulate a biological substrate for cellular receptor engagement and response. For this model system, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N- [6- [(2,4-dinitrophenyl)amino]hexanoyl] (ammonium salt) (DNP-cap-DPPE) was used for specific binding of anti-DNP IgE. IgE's bound to mast cell surface receptors (Fcis an element ofRI) aggregate and cluster over the patterned features if they are significantly smaller than the cells. For patterned features that are cell sized or larger, cells adhere to the silicon surface with receptors polarizing toward the edge of the patterned lipid. RBL cells with specifically engaged IgE-receptors are activated to undergo striking morphological changes that can be analyzed with fluorescence or scanning electron microscopy. This novel method for patterning antigen-functionalized lipids provides spatial control down to micron resolution for the antigenic stimulus while retaining the features of a plasma membrane substrate, including dynamics and variable composition. This method offers an alternative to microcontact printing with some enhanced capabilities for cellular immunological studies.