T Cell Modulation of Renal Ischemia Reperfusion Injury

描述（由申请人提供）：肾缺血再灌注损伤（IRI）是自体肾和同种异体肾中急性肾衰竭（ARF）的主要原因。我们和其他人已经证明了T细胞在实验性肾ARF中的重要调节作用。下一步是了解T细胞在肾IRI中的作用机制。我们假设T细胞通过运输到缺血性肾脏来调节肾脏IRI，通过T细胞受体（TCR）-主要组织相容性复合体（MHC）的抗原特异性相互作用被激活，并产生与其他特异性淋巴细胞协同工作的炎症介质。具体目标1：我们假设TCR-MHC复合物通过抗原特异性应答介导T细胞参与肾IRI。我们有初步的数据表明，TCR受体，以及MHC II类直接影响肾IRI的结果。我们将使用已建立的肾IRI小鼠模型和特异性基因敲除小鼠，并评估肾脏、分子和细胞反应。我们还将进行T细胞过继转移研究并使用骨髓嵌合体。我们将使用CD 4 TCR转基因小鼠来探讨抗原特异性和T细胞活化在IRI中的作用。具体目标2：我们假设早期T细胞运输，活化和细胞因子的产生介导肾IRI后的后续损伤。我们将使用我们新开发的淋巴细胞分离技术来评估T细胞进入缺血后肾脏的运输。我们将比较T细胞的运输与其他白细胞，并表征肾T细胞的激活状态和促炎细胞因子在蛋白质和mRNA水平的生产离体。连续转移技术将用于直接测试这些T细胞细胞因子的功能作用。具体目标3：我们假设IRI后，T细胞与NKT和B细胞相互作用以充分表达肾损伤。我们将使用NKT和B细胞功能缺陷的小鼠，并进行过继转移研究。我们将使用新开发的工具在野生型小鼠中进行激动剂、阻断剂和耗竭剂的补充实验，这些工具以前限制了这些研究。这是一个新的方向，将导致新的见解非常早期的细胞步骤，启动缺血后肾脏炎症。长期目标将是确定在肾IRI中促进损伤过程的抗原和分子途径，并开发针对ARF的靶向免疫疗法。

DESCRIPTION (provided by applicant): Kidney ischemia reperfusion injury (IRI) is a major cause of acute renal failure (ARF) in both native and allograft kidney. We and others have demonstrated an important modulatory role for T cells in experimental kidney ARF. The next stage is to understand the mechanisms underlying the role for T cells in renal IRI. We hypothesize that T cells modulate renal IRI by trafficking into ischemic kidney, becoming activated through antigen specific interactions through the T cell receptor (TCR)-major histocompatibility complex (MHC), and produce inflammatory mediators that work in concert with other specific lymphocytes. Specific Aim 1: We hypothesize that the TCR-MHC complex mediates T cell participation in renal IRI through antigen specific responses. We have preliminary data that TCR receptors, as well as MHC class II directly influence outcome of renal IRI. We will use an established mouse model of renal IRI and mice with specific knockouts, and evaluate renal, molecular and cellular responses. We will also perform T cell adoptive transfer studies and use bone marrow chimeras. We will use CD4 TCR transgenic mice to probe the role of antigen specificity and T cell activation in IRI. Specific Aim 2: We hypothesize that early T cell trafficking, activation and production of cytokines mediates subsequent injury following renal IRI. We will use our new developed lymphocyte isolation technique to evaluate trafficking of T cells into postischemic kidney. We will compare T cell trafficking with that of other leukocytes, and characterize the renal T cells ex vivo in terms of activation status and production of proinflammatory cytokines at the protein and mRNA level. Adoptive transfer techniques will be used to directly test the functional role of these T cell cytokines. Specific Aim 3: We hypothesize that T cells interact with NKT and B cells for full expression of renal injury after IRI. We will use mice deficient in NKT and B cell function, and perform adoptive transfer studies. We will perform complementary experiments with agonists, blockers and depleting agents in wild type mice using new developed tools that previously limited these studies. This is a novel direction that will lead to new insights into the very early cellular steps that initiate postischemic kidney inflammation. The long term goal will be to identify the antigens and molecular pathways that fuel the injury processes in renal IRI, and develop targeted immune therapies for ARF.