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Author:
Date:
2020-03-05
[Abstract] T cells are one major cell type of the immune system that use their T cell antigen receptor (TCR) to bind and respond to foreign molecules derived from pathogens. The ligand-TCR interaction half-lives determine stimulation outcome. Until recently, scientists relied on mutating either the TCR or its ligands to investigate how varying TCR-ligand interaction durations impacted on T cell activation. Our newly created opto-ligand-TCR system allowed us to precisely and reversibly control ligand binding to the TCR by light illumination. This system uses phytochrome B (PhyB) tetramers as a light-regulated TCR ligand. PhyB can be photoconverted between a binding (ON) and non-binding (OFF) conformation by 660 nm and 740 nm light illumination, respectively. PhyB ON is able to bind to a synthetic ...
[摘要] [摘要] T细胞是免疫系统的一种主要细胞类型，利用其T细胞抗原受体（TCR）结合并响应源自病原体的外来分子。配体-TCR相互作用的半衰期决定了刺激的结果。直到最近，科学家还是依靠突变TCR或其配体来研究变化的TCR-配体相互作用持续时间如何影响T细胞活化。我们新创建的光配体-TCR系统使我们能够通过光照精确且可逆地控制配体与TCR的结合。该系统使用植物色素B（PhyB ）四聚体作为光调节的TCR配体。PHYB 可光转化的结合（ON）之间，并且通过分别为660nm和740nm的光照射，非结合性（OFF）构象。PhyB ON能够结合通过将PhyB 相互作用因子（PIF）融合到TCRβ链而产生的合成TCR 。将PhyB 切换为OFF构象会破坏这种相互作用。PhyB 四聚体与PIF-TCR 足够长的结合导致T细胞活化（通过钙内流测量）。在这里，我们描述了有关如何为我们的光配体-TCR系统生成四聚体配体，如何通过流式细胞仪测量配体-TCR结合以及如何通过钙内流量化T细胞活化的协议。
[背景 ] ...
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