Maltz-Matyschsyk Michele, Melchiorre Clare K, Dostie-Laprise Kristen, Lynes Michael A
Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT 06269, United States.
Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT 06269, United States.
Cell Stress Chaperones. 2025 Jul 31;30(5):100106. doi: 10.1016/j.cstres.2025.100106.
Metallothionein (MT) is a highly conserved, low-molecular-weight (∼7 kDa), cysteine-thiol-rich, stress response protein essential to cellular homeostasis. Elevated MT levels can be induced in cells during response to oxidative stress, glucocorticoids, essential divalent cationic metals, toxic heavy metal cations, acute-phase cytokines, interferon-γ, and/or endotoxin exposure. MT isoforms 1 and 2 are expressed across most tissues/cells and are localized in cytosolic, nuclear, and extracellular environments, despite the absence of a signal peptide. Extracellular MT (eMT) plays a significant role in inflammatory disease by acting as a signal that modifies the functional profile of inflammatory cells. Treatment with anti-MT monoclonal antibody (UC1MT), which presumably targets the eMT, in various mouse models of inflammatory disease significantly reduces disease severity. This study examines the effects of eMT on T lymphocyte gene expression at exposure times of 5-90 min in vitro. Jurkat T-cells were treated with eMT alone or in combination with UC1MT, revealing distinct gene expression changes at all time points, with the most substantial effects observed at 90 min. The results demonstrated eMT's influence on G-protein-coupled receptor (GPCR) gene expression and cell proliferation, confirmed through calcium flux and Carboxyfluorescein Succinimdiyl Ester (CFSE) proliferation assays. An analysis at the 90-min time point identified a positive feedback loop wherein eMT induces additional MT messenger ribonucleic acid (mRNA) expression. Using an MT-GFP fusion vector, transfected Jurkat T-cells verified that eMT stimulates both MT transcript and protein expression. This study underscores eMT's role as an alarmin and its capacity to potentiate inflammatory disease by modulating gene and protein expression in T lymphocytes.
金属硫蛋白（MT）是一种高度保守的低分子量（约7 kDa）、富含半胱氨酸-硫醇的应激反应蛋白，对细胞内稳态至关重要。在细胞对氧化应激、糖皮质激素、必需二价阳离子金属、有毒重金属阳离子、急性期细胞因子、干扰素-γ和/或内毒素暴露的反应过程中，MT水平会升高。MT同工型1和2在大多数组织/细胞中均有表达，且尽管没有信号肽，但它们定位于胞质、核和细胞外环境中。细胞外MT（eMT）通过作为一种改变炎症细胞功能特征的信号，在炎症性疾病中发挥重要作用。在各种炎症性疾病小鼠模型中，用可能靶向eMT的抗MT单克隆抗体（UC1MT）进行治疗可显著降低疾病严重程度。本研究在体外5 - 90分钟的暴露时间内，考察了eMT对T淋巴细胞基因表达的影响。单独用eMT或与UC1MT联合处理Jurkat T细胞，发现在所有时间点均有明显的基因表达变化，在90分钟时观察到的影响最为显著。结果表明eMT对G蛋白偶联受体（GPCR）基因表达和细胞增殖有影响，这通过钙流和羧基荧光素琥珀酰亚胺酯（CFSE）增殖试验得到证实。在90分钟时间点的分析确定了一个正反馈回路，其中eMT诱导额外的MT信使核糖核酸（mRNA）表达。使用MT - GFP融合载体，转染的Jurkat T细胞证实eMT刺激MT转录本和蛋白表达。本研究强调了eMT作为警报素的作用及其通过调节T淋巴细胞中的基因和蛋白表达来增强炎症性疾病的能力。
