T-Lymphocyte role in Lung Ischemia-Reperfusion Injury

描述(逐字摘自申请人的摘要)克服 缺血再灌注损伤(I/R)是导致手术成功的关键事件 治疗性肺移植。我们的研究计划将检验整体 Th1-CD4+T淋巴细胞促进粒细胞介导的假说 大鼠肺缺血后微血管通透性增加 CD4O-CD4OL结合和趋化因子的产生。这一假设将得到检验。 通过旨在解决以下三个具体目标的试验： I)干扰CD4O-CD4OL相互作用减弱I/R诱导 微血管损伤；ii)白介素10(IL-10)，抑制Th1-CD4+ 淋巴细胞依赖的炎症反应，预防I/R诱导的肺损伤 通过调节Th1-CD4+淋巴细胞的黏附和激活及趋化因子 产生；iii)Th1-CD4+淋巴细胞/内皮细胞相互作用 肺泡后微循环导致I/R引起的通透性改变 这段血管。我们将测量循环中的淋巴细胞数量和 流式细胞术在分离大、小鼠淋巴细胞亚群中的应用 肺受到规定的I/R周期的影响。我们还将确定 大鼠和小鼠离体肺I/R微血管损伤的实验研究 微血管滤过系数(KFC)。KF关于Will的变化 用来确定CD4O-CD4OL信号在I/R损伤中的作用 使用针对这些分子中的每一种分子的抗体的过程 使用转基因小鼠。白介素10对小鼠肺损伤的保护作用 I/R引起的肺损伤将以类似的方式进行测试。评估 趋化因子的产生，特别是大鼠和小鼠肺中的MIP-2，将是 在所有实验中都进行了测试。最后，我们将确定是否 I/R诱导的肺泡后微循环内肯德基的增加是由于 淋巴细胞介导的炎症过程，使用节段性通透性 并通过使用免疫组织化学技术来识别 CD4O在福尔马林固定组织中的定位。这些研究的结果 将为淋巴细胞在急性I/R诱导中的作用提供新的见解 肺损伤，总体目标是提高临床肺治疗成功率 移植程序。

DESCRIPTION (Verbatim from the applicant's abstract) Overcoming ischemia-reperfusion injury (I/R) is a critical event leading to successful therapeutic lung transplantation. Our research proposal will test the overall hypothesis that Th1-CD4+ T-lymphocytes promote granulocytic-mediated microvascular permeability increase in the post-ischemic lung in part by CD4O-CD4OL binding and chemokine generation. The hypothesis will be tested through experimentation designed to address the following three specific aims: I) Interference with CD4O-CD4OL interaction attenuates I/R-induced microvasculai injury; II) Interleukin-10 (IL-10), which inhibits Th1 -CD4+ lymphocyte-dependent inflammatoryresponses, prevents I/R-induced lung injury via modulation of Th1 -CD4+ lymphocyte adhesion and activation and chemokine production; and III) Th1-CD4+ lymphocyte/ endothelial cell interactions in the post-alveolar microcirculation result in I/R induced permeability changes to this vascular segment. We will measure circulating lymphocyte numbers and differentiate lymphocyte subsets using flow cytometry in isolated rat and mouse lungs that are subjected to defined periods of I/R. We will also determine microvascular damage due to I/R in isolated rat and mouse lungs by measuring the microvascular filtration coefficient (Kfc). Changes in Kf about will then be used to determine the contribution of CD4O-CD4OL signaling in the I/R injury process using antibodies specific to each of these molecule' and through the use of genetically engineered mice. The protective effects of IL- 10 against I/R-induced lung damage will be tested in a similar manner. Assessment of chemokine production, specifically MIP-2 in the rat and mouse lung, will be performed in all experiments. Finally, we will determine whether the I/R-induced increase in Kfc within the post-alveolar microcirculation is due to lymphocytic-mediated inflammatory processes, using segmental permeability measures and by employing immunohistochemistry techniques for identifying the localization of CD4O in formalin-fixed tissues. The results of these studies will provide novel insight as to the role of lymphocytes in acute I/R-induced lung damage with the overall goal of improving success rates for clinical lung transplantation procedures.