IS AAA AN ANTIGEN-DRIVEN AUTOIMMUNE DISEASE? 'COLLABORAT

AAA的病因和发病机制尚不清楚。研究中需要验证的假设是抗原驱动的T细胞反应是否可能导致AAA的发生（并可能有助于繁殖），以及这些T细胞是否识别宿主抗原。我们的具体目标是：1。确定自发性或炎症性AAA患者的AAA病变中浸润的新鲜T细胞（未在培养中扩增）是否含有相当比例的单克隆T细胞。鉴定这些T细胞的TCR所使用的克隆扩增的α、β、γ和β链TCR转录本。2. 利用扩增的α链和β链TCR转录本在AAA炎性细胞浸润中鉴定T细胞识别的抗原。a.在AAA中表达克隆扩增的TCR转录本，通过转染将TCR转录本转入α β TCR阴性的α β J.RT3-T3.5 Jurkat细胞，或通过逆转录病毒载体感染正常的细胞毒性t淋巴细胞（CTL）系。b.确定这些转导或转染的T细胞是否具有克隆扩增的TCR，识别推定的AAA抗原（弹性蛋白，氧化LDL， AAA- p， I型和III型胶原，以及CMV）。3. 为了进一步确定整个AAA t细胞浸润的功能，并确保获得功能性应答的t细胞克隆，我们将通过限制稀释来开发针对假定的AAA抗原（弹性蛋白、氧化LDL、AAA- p /MAGP-36、I型和III型胶原以及CMV）特异性的t细胞克隆。a.充分表征这些t细胞克隆。b.比较这些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，以确定这些新鲜浸润T细胞的克隆群体是否与抗原特异性T细胞克隆具有相同的抗原特异性。4. 研究AAA病变（转录本和蛋白质）中趋化因子和Th1和Th2细胞因子的产生，并确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本交互式RO1计划第二次RO1研究拨款申请的结果联系起来，特别是与：(i)动脉瘤的形态和空间改变；（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.