利用供体猝灭荧光共振能量转移和荧光寿命成像荧光共振能量转移研究免疫突触形成过程中TCR-CD3链的动态质膜结合

Studying Dynamic Plasma Membrane Binding of TCR-CD3 Chains During Immunological Synapse Formation Using Donor-Quenching FRET and FLIM-FRET.

作者信息

Gagnon Etienne, Connolly Audrey, Dobbins Jessica, Wucherpfennig Kai W

机构信息

Department of Microbiology and Immunology, Institute for Research in Immunology and Cancer, Université de Montréal, PO Box 6128, Downtown Station, Montreal, Canada, H3C 3J7.

Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute, Boston, MA, 02115, USA.

出版信息

Methods Mol Biol. 2017;1584:259-289. doi: 10.1007/978-1-4939-6881-7_16.

DOI:10.1007/978-1-4939-6881-7_16

PMID:28255707

Abstract

Over the last decade, advancements in the time and space resolution of microscopy technologies have enabled dissection of the molecular events involved in T cell Immunological Synapse (IS) formation. Using a combination of Förster Resonance Energy Transfer (FRET) and Fluorescence Lifetime Imagining Microscopy (FLIM), we have demonstrated dynamic plasma membrane binding by cytoplasmic domains of T cell receptor (TCR)-associated CD3 chains and other T cell transmembrane receptors. We have developed methods for imaging such membrane binding both at steady state and during receptor triggering at the IS. Plasma membrane binding by cytoplasmic domains may represent a novel mechanism for regulating the signaling function of important receptors in the immune system.

摘要

在过去十年中，显微镜技术在时间和空间分辨率方面的进步使得对T细胞免疫突触（IS）形成过程中涉及的分子事件进行剖析成为可能。通过结合荧光共振能量转移（FRET）和荧光寿命成像显微镜（FLIM），我们已经证明了T细胞受体（TCR）相关CD3链的胞质结构域以及其他T细胞跨膜受体与动态质膜的结合。我们已经开发出在稳态以及IS处受体触发过程中对这种膜结合进行成像的方法。胞质结构域与质膜的结合可能代表了一种调节免疫系统中重要受体信号功能的新机制。