Roles of Activated Collagen V Stroma in Translant Rejection and Arteriopathies

我们已经报道了对胶原V [col（V）]的α 1（V）的自身免疫在急性和慢性炎症中的主要作用。 肺移植排斥反应这些研究的数据使我们假设，异常的alpha同源三聚体 1（V）链诱导抗col（V）自身免疫和器官移植排斥。为了验证这个假设，我们 研究了动脉粥样硬化的自身免疫成分是否涉及anficol（V）自身免疫， 仅仅基于在人动脉粥样硬化斑块中发现α 1（V）同源三聚体的事实。这里我们 证明人类和啮齿类动物的动脉粥样硬化实际上与anficol（V）自身免疫有关， 构成了支持α 1（V）同源三聚体形成之间联系的重要概念验证， 和抗col（V）自身免疫。我们提出的研究表明，异常的α 1（V）同源三聚体， 在闭塞性细支气管炎（OB）中表达，这是慢性肺移植排斥反应的基础; 同种异体移植物血管病（CAV），心脏移植中的动脉粥样硬化样排斥病理。我们还将 使用同源重组在体内有条件地诱导α 1（V）同源三聚体的表达， 小鼠成年肺上皮、成年平滑肌细胞和正常表达col（V）的所有成年组织中， 为了直接测试这种异常形式的col（V）在OB和CAV中的作用，在肺和心脏移植后， 在动脉粥样硬化中，当表达α 1（V）同源三聚体的小鼠与ApoE缺失的小鼠杂交时， 小鼠各种类型的免疫挑战和过继转移将进一步测试α 1（V）的作用 同源三聚体在这些新型小鼠模型系统中引发抗col（V）自身免疫中的作用。我们还将采用同源重组产生小鼠，其中通过肽分析发现最能被抗col（V）反应性T细胞和抗体识别的α 1（V）表位从体内α 1（V）基因中去除，以测试这些表位在OB、CAV、动脉粥样硬化和抗col（V）自身免疫中的真实作用。最后，我们提出了一系列体外和体内实验来测试“激活的”α 1（V）的概念， 包含可以增强和维持病理状态的基质，包括OB、CAV、动脉粥样硬化以及可能的其他病理，以及这样的基质如何影响细胞行为。

We've reported major roles for autoimmunity to the alpha 1 (V) of collagen V [col(V)] in acute and chronic lung transplant rejection. Data from these studies led us to hypothesize that abnormal homotrimers of alpha 1(V) chains induce anti-col(V) autoimmunity and organ transplant rejecfion. To test the hypothesis, we invesfigated whether the autoimmune component of atherosclerosis involves anfi-col(V) autoimmunity, based solely on the fact that alpha 1 (V) homotrimers are found in human atherosclerotic plaques. Here we demonstrate that atherosclerosis in humans and rodents is in fact associated with anfi-col(V) autoimmunity, constituting an important proof-of-concept in support ofthe link between alpha 1(V) homotrimer formafion and anti-col(V) autoimmunity. We propose studies to demonstrate that aberrent alpha 1(V) homotrimers are expressed in obliterative bronchiolitis (OB), which underiies chronic lung transplant rejection; and in cardiac allograft vasculopathy (CAV), an atherosclerosis-like rejection pathology in heart transplants. We will also use homologous recombinafion to condifionally induce expression of alpha 1(V) homotrimers in vivo in mouse adult lung epithelium, adult smooth muscle cells, and in ali adult tissues that normally express col(V) to directly test the roles of this aberrant form of col(V) in OB and CAV, upon lung and heart transplantation, respectively; and in atherosclerosis, upon crossing of alpha 1 (V) homotrimer-expressing mice with ApoE-null mice. Various types of immune challenges and adoptive transfers will further test the roles of alpha 1 (V) homotrimers in initiating anti-col(V) autoimmunity in these novel mouse model systems. We will also employ homologous recombination to generate mice in which alpha 1 (V) epitopes, found by peptide analysis to be the most recognized by anti-col(V) reactive T cells and antibodies, are removed from the alpha 1(V) gene in vivo, to test the true roles of such epitopes in OB, CAV, atherosclerosis and anti-col(V) autoimmunity. Finally, we propose a series of in vitro and in vivo experiments to test the concept of an "activated" alpha 1(V)- containing stroma that can enhance and perpetuate pathological states, including OB, CAV, atherosclerosis, and perhaps other pathologies as well, and how such a stroma affects cellular behaviors.