Redox control of CD8 T cell proliferation, differentiation and death

描述(申请人提供)：我们的目标是确定活性氧中间体(ROI)控制初始CD8T细胞激活、增殖和分化的分子机制。在幼稚的CD8T细胞激活过程中，酪氨酸磷酸化和钙内流迅速增加。与此同时，投资回报率也在增加。这种ROI的增加被认为是正常T细胞激活的关键，因为在病毒感染期间，与抗氧化剂共同孵育会减少CD8 T细胞在体外和体内的扩张。这一建议中提出的具体假设是，原始CD8 T细胞的激活、增殖和分化是由ROI通过控制信号蛋白中的半胱氨酸修饰为磺酸来控制的。特定目标1将测试调节ROI对初始CD8T细胞激活、增殖和分化的影响。将使用过表达过氧化氢酶和铜超氧化物歧化酶(CAT/SOD)或缺乏过氧化还蛋白II(PRX II-/-)的p14TCR转基因小鼠。从这些小鼠中提纯的初始CD8T细胞将在体外被多克隆刺激或与包被多肽的骨髓来源的树突状细胞相互作用而被激活。体内ROI调节对CD8T细胞活化、增殖和分化的影响将通过过继转移到未接种的受者体内，然后感染LCMV来确定。在特定目的2中，我们将确定蛋白酪氨酸磷酸酶SHP-1和SHP-2在CD8 T细胞激活、增殖和分化过程中的磺酸修饰。这将通过使用我们的新型标记试剂来实现，该试剂允许分离和鉴定含有磺酸的蛋白质。在特定目的2的第一部分，我们将确定这些PTPs在多克隆刺激激活的纯化的初始CD8T细胞中的修饰。在目标2的第二部分中，我们将确定包被多肽的树突状细胞激活的T细胞中PTPs是如何被修饰的。 公共卫生相关性：了解CD8 T细胞如何激活、增殖和分化，对于开发在疫苗接种或癌症治疗期间增加抗原特异性细胞数量的治疗方法至关重要，对于减少器官移植或自身免疫期间不需要的细胞也很重要。

DESCRIPTION (provided by applicant): Our goal is to determine the molecular mechanisms by which reactive oxygen intermediates (ROI) control the activation, proliferation and differentiation of naive CD8+ T cells. During naive CD8+ T cell activation there is a rapid increase in tyrosine phosphorylation and calcium influx. At the same time there is also an increase in ROI. This increase in ROI is thought to be critical to normal T cell activation as co-incubation with anti-oxidants decreases the expansion of CD8+ T cells in vitro and in vivo during viral infection. The specific hypothesis that is being addressed in this proposal is that naive CD8+ T cell activation, proliferation and differentiation are controlled by ROI through the controlled modification of cysteines in signaling proteins to sulfenic acid. Specific Aim 1 will test the effects of modulating ROI on naive CD8+ T cell activation, proliferation and differentiation. P14 TCR transgenic mice that overexpress both catalase and Cu superoxide dismutase (CAT/SOD) or lack peroxiredoxin II (Prx II -/-) will be used. Purified naive CD8+ T cells from these mice will be activated in vitro by polyclonal stimuli or after interaction with peptide coated bone marrow derived dendritic cells. The in vivo effects of ROI modulation on CD8+ T cell activation, proliferation and differentiation will be determined by adoptive transfer into naive recipients followed by LCMV infection. In Specific Aim 2 we will determine the sulfenic acid modification of the protein tyrosine phosphatases SHP-1 and SHP-2 during CD8+ T cell activation, proliferation and differentiation. This will be accomplished through the use of our novel labeling reagents which allow the isolation and identification of sulfenic acid containing proteins. In the first part of Specific Aim 2, we will determine the modification of these PTPs in purified naive CD8+ T cells activated by polyclonal stimuli. In the second part of aim 2, we will determine how PTPs are modified in T cells activated by peptide coated dendritic cells. PUBLIC HEALTH RELEVANCE: Understanding how CD8+ T cells become activated, proliferate and differentiate is critical to developing treatments to increase the number of antigen-specific cells during vaccination or cancer therapy and it is also important to reducing unneeded cells during organ transplant or autoimmunity.