Redox control of CD8 T cell proliferation, differentiation and death

描述（由申请人提供）：我们的目标是确定活性氧中间体（ROI）控制初始CD 8 + T细胞活化、增殖和分化的分子机制。在初始CD 8 + T细胞活化期间，酪氨酸磷酸化和钙内流快速增加。与此同时，ROI也在增加。ROI的这种增加被认为对正常T细胞活化至关重要，因为与抗氧化剂共孵育降低了病毒感染期间体外和体内CD 8 + T细胞的扩增。本提案中提出的具体假设是，初始CD 8 + T细胞活化、增殖和分化由ROI通过信号传导蛋白中的半胱氨酸受控修饰为次磺酸来控制。具体目标1将测试调节ROI对初始CD 8 + T细胞活化、增殖和分化的影响。将使用过表达过氧化氢酶和Cu超氧化物歧化酶（CAT/SOD）或缺乏过氧化物氧还蛋白II（Prx II -/-）的P14 TCR转基因小鼠。来自这些小鼠的纯化的初始CD 8 + T细胞将在体外通过多克隆刺激或在与肽包被的骨髓来源的树突细胞相互作用后活化。ROI调节对CD 8 + T细胞活化、增殖和分化的体内影响将通过过继转移到初始受体中，随后LCMV感染来确定。在具体目标2中，我们将确定在CD 8 + T细胞活化、增殖和分化过程中蛋白酪氨酸磷酸酶SHP-1和SHP-2的次磺酸修饰。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次磺酸的蛋白质。在第一部分的具体目标2，我们将确定这些PTP的修饰纯化的幼稚CD 8 + T细胞激活的多克隆刺激。在目标2的第二部分中，我们将确定PTPs如何在由肽包被的树突状细胞激活的T细胞中被修饰。 公共卫生相关性：了解CD 8 + T细胞如何被激活，增殖和分化对于开发治疗方法以增加疫苗接种或癌症治疗期间抗原特异性细胞的数量至关重要，并且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.