Targeting the inflammatory response to treat post-traumatic anxiety and depression.

部署的压力和暴露在创伤事件中使士兵面临比 公众对包括焦虑和抑郁在内的心理障碍的发展以及 创伤后应激障碍。这些精神障碍的发病率很高，而且 最近伊拉克和阿富汗战争(OEF/OIF)退伍军人的这些情况令人高度关注 对退伍军人管理局来说意义重大。虽然有许多治疗方案可用于治疗这些疾病 在某些情况下，他们对经典的抗焦虑和抗抑郁药物治疗有轻微反应 是创伤应激暴露的结果。对……领域的广泛研究 精神神经免疫学表明，这些情况与免疫调节失调有关。 功能，表现为全身炎症增加，细胞和体液免疫改变； 被认为是这些疾病的病理生理学基础的机制。我们以前的研究和 文献中的其他研究表明，T细胞对创伤应激暴露有反应，并能影响 小鼠的行为反应，根据应激的类型赋予其韧性或敏感性 T细胞和细胞因子环境。我们的初步结果有力地表明CD8+细胞毒性T细胞是主要的 全身和脑部炎症的来源，并促进发展适应不良行为的易感性 对压力的反应。另一方面，我们最近发表的研究表明，CD4+T细胞 对压力的行为反应，可能是通过减少炎症，与该领域的其他人的工作一致。 因此，这一应用的总体目标是在临床前小鼠模型中测试治疗效果 通过减少由创伤应激暴露引发的炎症过程来治疗焦虑和抑郁。 我们建议特异性地操纵CD4+和CD8+T细胞介导的免疫，以减少全身和大脑 炎症，并改善行为结果。我们提出了三个具体目标： 具体目标1将通过检查确定创伤应激改变T细胞功能的机制 CD4+和CD8+T细胞在应激反应中的归巢特性，以及它们是否在 糖皮质激素受体敏感性。为了实现这一点，我们将用T细胞重建T细胞缺陷的Rag2-/-小鼠 来自表达绿色荧光蛋白(GFP)的小鼠的细胞亚群，允许跟踪和 识别包括大脑在内的多个组织中的T细胞。此外，我们还将进行转录组 应激与非应激野生型小鼠T细胞的RNA测序分析 创伤应激诱导T细胞活化的途径。具体目标2旨在确定影响 应激小鼠操纵CD4+和CD8+T细胞行为的研究该方法将涉及一个) 应激野生型小鼠的CD4+或CD8+T细胞在Rag2-/-小鼠中的重建，以及b)使用 应激野生型小鼠的抗CD4+或CD8+T细胞中和抗体。经过治疗后，小鼠将 被评估为焦虑、行为绝望和惊吓反应，作为情绪行为的衡量标准。特定的 目的3将研究操纵CD4+和CD8+T细胞对外周和脑部炎症的影响。 将评估外周组织和大脑中的血浆和组织细胞因子水平；神经炎症将被 用小胶质细胞培养和流式细胞仪进一步评估。最后，为了确定CD8+T细胞是否赋予其 通过细胞因子肿瘤坏死因子-α和白介素6的作用发挥作用。我们还将阻止这些细胞因子的存在 检测CD8+T细胞的数量，并确定是否可以减少神经炎症。 这项应用的研究有望为CD8+T细胞是主要的概念提供证据 创伤应激暴露引发炎症过程的来源，并有可能改善 以这些细胞为靶点进行情绪调节。此外，它们可能有助于识别压力的独特机制。 诱导T细胞活化和治疗应激相关精神障碍的新靶点。

The stress of deployment and exposure to traumatic events puts soldiers at a greater risk than the general public for the development of psychological disorders, including anxiety and depression, as well as post-traumatic stress disorder. These mental disorders occur with high comorbidity, and the prevalence of these conditions in Veterans of the recent Iraq and Afghanistan wars (OEF/OIF) is a concern of high significance for the VA. While a number of therapeutic options are available for the treatment of these conditions, they are marginally responsive to classical anxiolytic and antidepressant treatment when they develop as a consequence of traumatic stress exposure. Extensive research in the field of psychoneuroimmunology has indicated that these conditions are associated with dysregulated immune function, manifested as increased systemic inflammation and altered cellular and humoral immunity; which is believed to be a mechanism underlying the pathophysiology of these disorders. Our previous studies and others in the literature have shown that T cells are responsive to traumatic stress exposure and can influence behavioral responses of mice, conferring either resilience or susceptibility to stress depending upon the type of T cell and the cytokine milieu. Our preliminary results strongly indicate that CD8+ cytotoxic T cells are a major source of systemic and brain inflammation and promote susceptibility to develop maladaptive behavioral responses to stress. On the other side, our recently published study indicates that CD4+ T cells improve behavioral responses to stress, perhaps by reducing inflammation, in line with the work of others in the field. Thus, the overall objective of this application is to test, in a pre-clinical mouse model, the therapeutic efficacy of treating anxiety and depression by reducing inflammatory processes triggered by traumatic stress exposure. We propose to specifically manipulate CD4+ and CD8+ T cell mediated immunity to reduce systemic and brain inflammation, and improve behavioral outcomes. We propose 3 specific aims: Specific Aim 1 will identify mechanisms by which traumatic stress alters T cell functions by examining homing properties of CD4+ and CD8+ T cells in response to stress, and whether they develop alterations in glucocorticoid receptor sensitivity. To accomplish this we will reconstitute T cell deficient Rag2-/- mice with T cell subsets derived from green fluorescent protein (GFP) expressing mice, allowing for the tracking and identification of T cells in multiple tissues- including the brain. Additionally, we will conduct transcriptome analysis of T cells of stressed vs non-stressed wild type mice using RNA-sequencing (RNAseq) to identify pathways of T cell activation induced by traumatic stress. Specific Aim 2 is designed to determine the effects on behavior of manipulating CD4+ and CD8+ T cells in stressed mice. The approach will involve a) reconstitution in Rag2-/- mice with CD4+ or CD8+ T cells from stressed wild type mice, and b) the use of neutralizing antibodies against CD4+ or CD8+ T cells in stressed wild type mice. Following treatment, mice will be assessed for anxiety, behavioral despair, and startle reactivity as measures of emotional behavior. Specific Aim 3 will study the effects of manipulating CD4+ and CD8+ T cells on peripheral and brain inflammation. Plasma and tissue cytokine levels will be evaluated in peripheral tissue and brain; neuroinflammation will be further assessed using microglial cultures and flow cytometry. Finally, to determine if CD8+ T cells confer their effects through the actions of the cytokines TNF-α and IL-6. We will also block these cytokines in the presence of CD8+ T cells and determine if there is a reduction in neuroinflammation. The studies in this application are expected to provide proof of concept that CD8+ T cells are the main source of inflammatory processes triggered by traumatic stress exposure and it is possible to improve emotional regulation by targeting these cells. Furthermore, they may help identify unique mechanisms of stress induced T cell activation and novel targets of therapeutic intervention to treat stress related mental disorders.