A translational model of neuro-immune therapy for PTSD in veterans of the OEF/OIF

描述（由申请人提供）： 创伤后应激障碍（PTSD）是最常见的战争相关疾病之一，并与其他焦虑症高度共病。最近的伊拉克和阿富汗战争（持久自由行动/伊拉克自由行动）的退伍军人患有创伤后应激障碍和其他精神障碍，如焦虑和情绪障碍的发病率特别高。治疗PTSD的一个重要问题是缺乏适当的药理学或生物学干预来治疗这种疾病。事实上，目前用于PTSD的药物用于治疗相关症状，如抑郁症或睡眠问题，并且没有有效治疗PTSD的制剂。 最近的研究表明，T淋巴细胞对创伤后应激和焦虑的发展具有保护作用，正如小鼠研究所表明的那样。这些研究表明，增强T细胞介导的反应赋予保护，防止急性暴露于捕食者气味后对压力的焦虑和惊吓反应的发展。这些研究进一步表明，缺乏T细胞反应的小鼠更容易受到压力的影响。这与我们的初步结果一致，表明其他T细胞缺乏的小鼠模型在急性应激暴露后更容易发生焦虑，并且用来自野生型的CD 4 + T细胞重建恢复了对应激的适应性反应。本申请的目的是验证和建立焦虑和应激相关障碍中T细胞功能的小鼠模型，以研究T细胞在赋予针对创伤后应激和焦虑的保护中的作用。核心假设是T细胞将通过运输到大脑中刺激神经营养因子的产生来保护心因性应激。进一步假设极化的辅助性T细胞2型（TH 2）将介导这些效应，而TH 1将有利于病理性应激反应的发展。为了进行测试，我们将在2种PTSD行为模型中使用缺乏功能性T和B细胞的RAG 2-/-缺陷小鼠，以及通过过继转移从BALB/c野生型小鼠重建的T细胞。T细胞的体外分化将被应用于测试TH 2与TH 1淋巴细胞对创伤后应激和焦虑的作用。用分化的T细胞重建的BALB/c野生型、RAG 2-/-和RAG 2-/-小鼠将在暴露于捕食者气味范例中或在条件巴甫洛夫恐惧反应的消退中进行评估。第一种范式模拟了急性暴露于心因性应激以及随后对应激和焦虑的反应，第二种范式模拟了适应不良的恐惧反应。这两种模式模拟了创伤后应激障碍中受影响的大脑功能的关键方面。实时RT-PCR将用于比较脑中细胞因子mRNA的表达，以评价脑炎症反应和T细胞治疗的效果。将这些应答与通过ELISA测定的细胞因子的外周水平进行比较。免疫组织化学和共聚焦显微镜将被用来分析T细胞在大脑中的存在和神经营养因子的表达。在具体目标#1中，我们将比较BALB/c野生型和RAG 2-/-小鼠之间的行为、免疫学和神经化学反应。在具体目标#2中，我们将测试用分化的TH 2细胞重建RAG 2-/-小鼠的效果，在具体目标#3中，我们将测试用分化的TH 1细胞重建RAG 2-/-小鼠在行为、免疫和神经化学反应中的效果。 本申请中提出的实验和研究旨在建立PTSD中神经免疫功能的临床前模型，并测试分化的T细胞在心因性创伤应激和焦虑中的作用。这些研究可以提供对心因性压力恢复力的细胞机制的深入了解，当受到刺激时，可以在大脑中提供再生和修复功能，并恢复正常的压力反应和行为。这将有助于制定新的和更有效的干预战略，以治疗创伤后应激障碍和焦虑相关疾病。 公共卫生相关性： 创伤后应激障碍（PTSD）在伊拉克和阿富汗战争地区的退伍军人中非常普遍。目前使用精神药物的药物治疗显示出较差的疗效，并且通常旨在治疗相关症状而不是疾病。目前的研究将评估T细胞介导的过程是否可以保护创伤后应激和焦虑的发展。假设TH 2细胞在暴露于急性应激后将运输到大脑，在那里它们将刺激神经营养因子的释放，从而保护免受应激的有害影响。这些研究是基于最近的发现，表明T细胞可以防止PTSD小鼠模型中焦虑和过度应激反应的发展。这些研究旨在建立一种动物模型，以测试T细胞治疗恢复正常应激反应和预防创伤后应激障碍和焦虑症的疗效。

DESCRIPTION (provided by applicant): Posttraumatic stress disorder (PTSD) is one of the most common war-related illness and is highly comorbid with other anxiety disorders. Veterans from the recent Iraq and Afghanistan wars (Operation Enduring Freedom/Operation Iraqi Freedom) have a particularly high incidence of PTSD and other mental disorders such as anxiety and mood disorders. A significant problem for the treatment of PTSD is the lack of an appropriate pharmacological or biological intervention to treat the illness. Indeed, current medications for PTSD are used to treat associated symptoms such as depression or sleep problems and there in no formulation that effectively treat PTSD. Recent studies indicate that T lymphocytes are protective against the development of posttraumatic stress and anxiety as suggested by studies in mice. These studies show that boosting T cell mediated responses confer protection against the development of anxiety and startle responses to stress after acute exposure to predator odor. These studies further indicate that mice deficient of T cell responses are more susceptible to the effects of stress. This is in agreement with our preliminary results suggesting that other mice models of T cell deficiency are more susceptible to the development of anxiety after acute stress exposure and that reconstitution with CD4+ T cells from wild type restore adaptive responses to stress. The objective of the present application is to validate and establish a mice model of T cell function in anxiety and stress-related disorders to study the role of T cells in conferring protection against posttraumatic stress and anxiety. The central hypothesis is that T cells will confer protection to psychogenic stress by trafficking into the brain where they will stimulate the production of neurotrophic factors. It is further hypothesized that polarized T helper type 2 (TH2) cells will mediate these effects while TH1 will favor the development of pathological stress responses. To test, we will employ the RAG2-/- deficient mice which lack functional T and B cells and reconstitution of T cells by adoptive transfer from BALB/c wild type mice in 2 behavioral models of PTSD. In vitro differentiation of T cells will be applied to test the role of TH2 vs TH1 lymphocytes on posttraumatic stress and anxiety. BALB/c wild type, RAG2-/- and RAG2-/- mice reconstituted with differentiated T cells will be evaluated in the exposure to predator odor paradigm or in the extinction of conditioned Pavlovian fear responses. The first paradigm models acute exposure to psychogenic stress and later responses to stress and anxiety and the second paradigm models maladaptive fear responses. These 2 paradigms model key aspects of brain function which are affected in PTSD. Real-time RT-PCR will be used to compare cytokine mRNA expression in the brain to evaluate brain inflammatory responses and the effects of T cell treatment. These responses will be compared with peripheral levels of cytokines determined by ELISA. Immunohistochemistry and confocal microscopy will be employed to analyze the presence of T cells in the brain and the expression of neurotrophic factors. In specific aim #1we will compare behavioral, immunological and neurochemical responses between BALB/c wild type and RAG2-/- mice. In specific aim #2 we will test the effects of reconstituting RAG2-/- mice with differentiated TH2 cells and in specific aim #3 we will test the effects of reconstituting RAG2-/- mice with differentiated TH1 cells in behavioral, immunological and neurochemical responses. The experiments and studies proposed in this application are aimed at establishing a pre-clinical model of neuroimmune function in PTSD and to test the role of differentiated T cells in psychogenic traumatic stress and anxiety. These studies may provide insight into cellular mechanisms of resilience to psychogenic stress that when stimulated may provide regenerative and repair functions in the brain and restore normal stress responses and behavior. This will assist in the development of new and more effective strategies of intervention for the treatment of PTSD and anxiety-related disorders. PUBLIC HEALTH RELEVANCE: Posttraumatic stress disorders (PTSD) are highly prevalent in veterans of the Iraq and Afghanistan war zones. Current pharmacological treatments with psychotropic medications display poor efficacy and are generally aimed at treating associated symptoms rather than the illness. The present studies will evaluate if processes mediated by T cells confer protection against the development of posttraumatic stress and anxiety. It is hypothesized that TH2 cells will traffic to the brain after exposure to acute stress where they will stimulate the release of neurotrophic factors conferring protection against deleterious effects of stress. These studies are based upon recent findings suggesting that T cells may prevent the development of anxiety and exaggerated stress responses in a mice model of PTSD. These studies are aimed at establishing an animal model to test the efficacy of treatments with T cells to restore normal stress responses and protect against PTSD and anxiety disorders.