Sensitivity of TCR signaling to GSH loss in HIV disease

描述（由申请人提供）：先前在我们实验室和其他地方的工作已经确定，根据细胞内谷胱甘肽（GSH）水平测量的细胞内氧化还原状态调节T细胞功能。炎症反应如NF-kappaB的激活在氧化应激下（当GSH耗尽时）被上调，而T细胞（TCR）信号所需的激活事件，包括酪氨酸激酶磷酸化和钙内流，在这些条件下被下调，但在细胞内GSH水平正常时则被下调。与这些发现一致的是，细胞内谷胱甘肽在多种T细胞功能受损的疾病（如HIV感染、败血症、糖尿病、肿瘤）中往往会被耗尽。

DESCRIPTION (provided by applicant): Previous work in our laboratory and elsewhere has established that intracellular redox status measured in terms of intracellular glutathione (GSH) levels regulates T cell function. Inflammatory responses such as the activation of NF-kappaB are up-regulated under oxidative stress (when GSH is depleted) whereas activation events required for T cell (TCR) signaling, including tyrosine kinase phosphorylations and calcium influx, are down-regulated under these conditions but favored when intracellular GSH levels are normal. Consistent with these findings, intracellular GSH tends to be depleted in a wide variety of diseases (e.g., HIV infection, sepsis, diabetes, neoplasia) in which T cell function is often impaired. To define the biochemical mechanisms that underlie the redox regulation of TCR signaling, we propose here to combine the 11-color Fluorescent Activated Cell Sorter (Hi-D FACS) technology that we have developed to measure intracellular redox status in human T cell subsets with Hi-D FACS methods to quantitate phosphorylation of individual kinases in these T cell subsets. By examining the functional consequences of altering redox status (depleting/repleting GSH) in freshly isolated T cells from healthy donors, and by probing the relationship between redox status and T cell function in patients with various diseases, we contribute significantly to basic and clinical understanding of the impact of GSH depletion on immune function. Further, since treatment with non-toxic cysteine pro-drugs can replenish GSH, our findings may open the way to introduction of adjunct therapies to increase immune surveillance and bolster responses to infection. Finally, the technology we develop here will provide new and better ways to monitor disease progress and evaluate therapeutic interventions.