IS AAA AN ANTIGEN-DRIVEN AUTOIMMUNE DISEASE? 'COLLABORAT

AAA的病因和发病机制尚不清楚。将在GRANT中检验的假设是，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AAA的繁殖)，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AAA皮损中的新鲜(未扩增)T细胞是否含有相当比例的单克隆性T细胞。以确定这些T细胞的TCR所使用的克隆性扩展的α、β、伽马和β链TCR转录本。2.利用克隆性扩增的α链和β链TCR转录本鉴定AAA炎性细胞浸润物中T细胞识别的抗原。A.为了在AAA中表达克隆扩增的TCR，通过转染将TCR转录本渗透到αβTCR阴性的αβJRT3-T3.5 Jurkat细胞，或通过感染逆转录病毒载体进入正常的细胞毒性T淋巴细胞(CTL)系。B.确定这些T细胞是否转导或转染了克隆扩增的TCR，识别可能的AAA抗原(弹性蛋白、氧化型低密度脂蛋白、AAA-P、I型和III型胶原以及CMV)。3.为了进一步确定整个AAA T细胞的功能，并确保获得有功能反应的T细胞克隆，我们将通过限制稀释法开发针对AAA抗原(S)(弹性蛋白、氧化型低密度脂蛋白、AAA-P/MAGP-36、I型和III型胶原以及巨细胞病毒)的T细胞克隆。A.充分描述这些T细胞克隆的特征。B.比较这些T细胞克隆所使用的TCR序列与来自同一患者的新鲜(未经培养扩增)AAA浸润性T细胞的TCR序列，以确定这些新鲜浸润性T细胞的克隆群是否具有与抗原特异性T细胞克隆相同的抗原特异性。4.研究AAA病变(转录本和蛋白质)中趋化因子和Th1、Th2细胞因子的产生，并鉴定产生这些分子的单个细胞类型。为了将AAA病变的浸润程度、浸润性T细胞的激活阶段、T细胞的寡克隆性群体的存在以及趋化因子和细胞因子的产生与第二个RO1研究资助项目的应用相关联，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.