IS AAA AN ANTIGEN-DRIVEN AUTOIMMUNE DISEASE? 'COLLABORAT

AAA 的病因和发病机制尚不清楚。 待验证的假设是抗原驱动的 T 细胞反应是否可能导致 AAA 的启动（并可能有助于传播），以及这些 T 细胞是否识别宿主抗原。 我们的具体目标是： 1. 确定浸润自发性或炎性 AAA 患者 AAA 病变的新鲜（未在培养中扩增）T 细胞是否含有相当比例的单克隆 T 细胞。 鉴定这些 T 细胞的 TCR 使用的克隆扩增的 α、β、γ 和 β 链 TCR 转录本。 2. 利用 AAA 炎症细胞浸润中克隆扩增的 α 链和 β 链 TCR 转录物来鉴定 T 细胞识别的抗原。 一个。为了表达在 AAA 中克隆扩增的 TCR 转录物，通过转染将 TCR 转录物渗透到 alpha beta TCR 阴性的 alpha beta J.RT3-T3.5 Jurkat 细胞中，或通过用逆转录病毒载体感染来渗透到正常细胞毒性 T 淋巴细胞 (CTL) 系中。 b.为了确定这些用克隆扩增的 TCR 转导或转染的 T 细胞是否识别假定的 AAA 抗原（弹性蛋白、氧化 LDL、AAA-P、I 型和 III 型胶原蛋白以及 CMV）。 3. 为了进一步定义整个 AAA T 细胞浸润的功能并确保获得功能响应的 T 细胞克隆，我们将通过有限稀释开发对推定 AAA 抗原（弹性蛋白、氧化 LDL、AAA-P/MAGP-36、I 型和 III 型胶原蛋白以及 CMV）具有特异性的 T 细胞克隆。 一个。全面表征这些 T 细胞克隆。 b.将这些 T 细胞克隆使用的 TCR 序列与来自同一患者的新鲜（未在培养中扩增）AAA 浸润 T 细胞的 TCR 序列进行比较，以确定这些新鲜浸润 T 细胞的克隆群是否与抗原特异性 T 细胞克隆具有相同的抗原特异性。 4. 研究 AAA 病变中趋化因子以及 Th1 和 Th2 细胞因子的产生（转录本和蛋白质），并鉴定产生这些分子的单个细胞类型。 为了将浸润程度、浸润 T 细胞的激活阶段、T 细胞寡克隆群体的存在以及 AAA 病变中趋化因子和细胞因子的产生与该交互式 RO1 计划的第二个 RO1 研究资助申请的结果联系起来，特别是与： (i) 动脉瘤的形态和空间变化； (ii) 动脉瘤的生物力学特性。

The etiology and pathogenesis of AAA is poorly understood. The hypothesis to be tested in grant is whether an antigen-driven T-cell response may be responsible for the initiation (and may contribute to the propagation) of AAA, and whether these T cells recognize host antigens. Our specific aims are: 1. To determine whether fresh (not expanded in culture) T cells infiltrating AAA lesions of patients with spontaneous or inflammatory AAA, contain substantial proportions of monoclonal T cells. To identify clonally expanded alpha-, beta-, gamma- and beta-chain TCR transcripts employed by the TCRs of these T cells. 2. To identify the antigens recognized by T cells employing the clonally expanded alpha- and beta-chain TCR transcripts in AAA inflammatory cell infiltrates. a. To express the clonally expanded in AAA infiltrates TCR transcripts into alpha beta TCR-negative into alpha beta J.RT3-T3.5 Jurkat cells by transfection, or into normal cytotoxic T-lymphocyte (CTL) lines by infection with retroviral vectors. b. To determine whether these transduced or transfected T cells with the clonally expanded TCR, recognize putative AAA antigens (elastin, oxidized LDL, AAA-P, collagen types I and III, and CMV). 3. To further define the functionality of the entire AAA T-cell infiltrate and to ensure that functionally responsive T-cell clones are obtained, we will develop by limiting dilution T-cell clones specific for putative AAA antigen(s) (elastin, oxidized LDL, AAA-P/MAGP-36, collagen types I and III, and CMV). a. To fully characterize these T-cell clones. b. To compare the TCR sequences used by these T-cell clones to those of fresh (not expanded in culture) AAA infiltrating T cells from the same patients, in order to determine whether clonal populations of these fresh infiltrating T cells have the same antigenic specificities to those of antigen-specific T-cell clones. 4. To investigate the production of chemokines and Th1 and Th2 cytokines in AAA lesions (transcripts and proteins) and to identify the individual cell types producing these molecules. To correlate the degree of infiltration, the activation stage of the infiltrating T cells, the presence of oligoclonal populations of T cells and the production of chemokines and cytokines in AAA lesions with the findings of the second RO1 research grant application of this Interactive RO1 Program and, in particular, with: (i) Morphological and spatial alterations in the aneurysm; (ii) Biomechanical properties of the aneurysm.