ACS Appl Mater Interfaces. 2018 Apr 11;10(14):11486-11494. doi: 10.1021/acsami.7b19643. Epub 2018 Mar 30.
Studying how different signaling pathways spatially integrate in cells requires selective manipulation and control of different transmembrane ligand-receptor pairs at the same time. This work explores a novel method for precisely arranging two arbitrarily chosen ligands on a micron-scale two-dimensional pattern. The approach is based on lithographic patterning of Au and TiO films, followed by their selective functionalization with Ni-nitrilotriacetic acid-histidine and biotin-avidin chemistries, respectively. The selectivity of chemical and biological functionalizations is demonstrated by X-ray photoelectron spectroscopy and immunofluorescence imaging, respectively. This approach is applied to produce the first type of bifunctional surfaces with controllably positioned ligands for activating the receptors of natural killer (NK) immune cells. NK cells were used as a model system to demonstrate the potency of the surface in guiding site-selective cell attachment and activation. Upon applying the suitable ligand or ligand combination, the surfaces guided the appropriate single- or bifunctional attachment and activation. These encouraging results demonstrate the effectiveness of the system as an experimental platform aimed at the comprehensive understanding of the immunological synapse. The great simplicity, modularity, and specificity of this approach make it applicable for a myriad of combinations of other biomolecules and applications, turning it into the "Swiss knife" of biointerfaces.
研究不同信号通路如何在细胞中空间整合，需要同时对不同的跨膜配体-受体对进行选择性操作和控制。本工作探索了一种在微米尺度二维图案上精确排列任意两个配体的新方法。该方法基于 Au 和 TiO 薄膜的光刻图案化，随后分别通过 Ni- 氮基三乙酸-组氨酸和生物素-亲和素化学进行选择性功能化。化学和生物功能化的选择性分别通过 X 射线光电子能谱和免疫荧光成像来证明。该方法应用于产生具有可控定位配体的第一类双功能表面，以激活自然杀伤 (NK) 免疫细胞的受体。NK 细胞被用作模型系统，以证明表面在引导位点选择性细胞附着和激活方面的功效。通过施加合适的配体或配体组合，表面引导适当的单功能或双功能附着和激活。这些令人鼓舞的结果表明，该系统作为一个实验平台，可用于全面了解免疫突触。该方法具有极大的简单性、模块化和特异性，适用于其他生物分子和应用的无数组合，使其成为生物界面的“瑞士军刀”。
