■CD8+细胞毒性T淋巴细胞(CTL)在防御病毒感染和肿瘤方面非常有效。它们通过T细胞受体(TCR)识别肽-MHC-I复合物和共刺激而被激活。这种同源相互作用促进涉及细胞骨架重排的紧密细胞-细胞连接的组织,以实现效应子功能和靶细胞的清除。这是裂解颗粒不对称运输和动员到细胞-细胞接触的关键,促进裂解介质如颗粒酶和穿孔素的定向分泌。线粒体通过控制钙通量等过程在调节CTL功能中发挥作用,通过氧化磷酸化提供必要的能量,以及它自己在70S核糖体上的蛋白质翻译。然而,在成熟的CTL中尚未研究过急性抑制细胞溶质翻译在TCR后的快速反应中的作用。
■这里,我们研究了早期TCR激活和CD28共刺激对细胞骨架动态重组后人类CTL中胞质蛋白合成的重要性,线粒体,和裂解颗粒通过环己酰亚胺对80S核糖体的短期化学抑制和嘌呤霉素对80S和70S的短期化学抑制。
■我们观察到真核核糖体功能是允许微管蛋白细胞骨架和线粒体的适当不对称重组以及在人原代CTL中TCR激活后的早期mTOR途径激活所必需的。
■胞质蛋白翻译是在TCR激活时增加葡萄糖代谢和脱粒能力并因此调节人CTL的全部效应子功能所必需的。
UNASSIGNED: CD8+ cytotoxic T lymphocytes (CTLs) are highly effective in defending against viral infections and tumours. They are activated through the recognition of peptide-MHC-I complex by the T-cell receptor (TCR) and co-stimulation. This cognate interaction promotes the organisation of intimate cell-cell connections that involve cytoskeleton rearrangement to enable effector function and clearance of the target cell. This is key for the asymmetric transport and mobilisation of lytic granules to the cell-cell contact, promoting directed secretion of lytic mediators such as granzymes and perforin. Mitochondria play a role in regulating CTL function by controlling processes such as calcium flux, providing the necessary energy through oxidative phosphorylation, and its own protein translation on 70S ribosomes. However, the effect of acute inhibition of cytosolic translation in the rapid response after TCR has not been studied in mature CTLs.
UNASSIGNED: Here, we investigated the importance of cytosolic protein synthesis in human CTLs after early TCR activation and CD28 co-stimulation for the dynamic reorganisation of the cytoskeleton, mitochondria, and lytic granules through short-term chemical inhibition of 80S ribosomes by cycloheximide and 80S and 70S by puromycin.
UNASSIGNED: We observed that eukaryotic ribosome function is required to allow proper asymmetric reorganisation of the tubulin cytoskeleton and mitochondria and mTOR pathway activation early upon TCR activation in human primary CTLs.
UNASSIGNED: Cytosolic protein translation is required to increase glucose metabolism and degranulation capacity upon TCR activation and thus to regulate the full effector function of human CTLs.