Use of TIRF to Monitor T-Lymphocyte Membrane Dynamics with Submicrometer and Subsecond Resolution.
Use of TIRF to Monitor T-Lymphocyte Membrane Dynamics with Submicrometer and Subsecond Resolution.
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DOI：
10.1007/s12195-014-0361-8
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发表时间：
2015
影响因子：
2.8
通讯作者：
Pierres, Anne
中科院分区：
文献类型：
--
作者：
Brodovitch, Alexandre;Limozin, Laurent;Bongrand, Pierre;Pierres, Anne
关键词：
获得性免疫反应的一个关键步骤是T淋巴细胞在持续几分钟的接触中快速和特异地检测特定细胞上是否存在外来抗原的能力。许多证据表明，T细胞受体和配体之间的分子相互作用的寿命与T细胞的激活之间存在着深刻的联系，但键的形成和解离的确切机制仍不完全清楚。以前用干涉反射显微镜/反射干涉对比显微镜所做的实验显示，微米尺寸的膜区的横向运动具有几个纳米的平均幅度。最近，全内反射荧光显微镜被用来显示初级T淋巴细胞与模型表面之间的初始相互作用涉及微绒毛的尖端(通常为0.2微米平方米的面积)，产生几秒钟的明显接触，使细胞能够检测到其膜受体的配体。在这里，我们发现这些微绒毛显示出最小的侧向位移，但定量荧光测量表明，在1-S观测期间，发生了幅度为67 nm的自发横向波动。这可能在膜受体参与和随后的信号产生中发挥重要作用。
A key step of adaptive immune responses is the T lymphocyte capacity to detect the presence of foreign antigens on specialized cells with high speed and specificity during contacts lasting a few minutes. Much evidence suggests that there is a deep link between the lifetime of molecular interactions between T cell receptors and ligands and T cell activation, but the precise mechanisms of bond formation and dissociation remain incompletely understood. Previous experiments done with interference reflection microscopy/reflection interference contrast microscopy disclosed transverse motions with several nanometer average amplitude of micrometer size membrane zones. More recently, total internal reflection fluorescence microscopy was used to show that the initial interaction between primary T lymphocytes and model surfaces involved the tip of microvilli (typically 0.2 µm2 area) generating apparent contacts of a few seconds that allowed cells to detect ligands of their membrane receptors. Here we show that these microvilli displayed minimal lateral displacements but quantitative fluorescence measurement suggested the occurrence of spontaneous transverse fluctuations of order of 67 nm amplitude during 1-s observation periods. This may play a major role in membrane receptor engagement and ensuing signal generation.
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