IS AAA AN ANTIGEN-DRIVEN AUTOIMMUNE DISEASE? 'COLLABORAT

AAA 是一种抗原驱动的自身免疫性疾病吗？

• 批准号: 6527327
• 负责人: Chris D Platsoucas
• 金额: $ 33.75万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6527327
• 关键词: T cell receptor; T lymphocyte; abdomen; aorta aneurysm; autoantigens; autoimmune disorder; chemokine; cooperative study; cytokine; disease /disorder etiology; extracellular matrix proteins; lymphocyte proliferation; tissue /cell culture; transfection

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.

AAA的病因和发病机制知之甚少。 在Grant中要检验的假设是抗原驱动的T细胞应答是否可能导致AAA的起始（并且可能有助于AAA的传播），以及这些T细胞是否识别宿主抗原。 我们的具体目标是：1.确定自发性或炎性AAA患者的AAA病变浸润的新鲜（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所使用的克隆扩增的α-、β-、γ-和β-链TCR转录物。 2.在AAA炎性细胞浸润中采用克隆扩增的α和β链TCR转录物鉴定T细胞识别的抗原。 a.通过转染将在AAA浸润细胞中克隆扩增的TCR转录物表达为α β TCR阴性的α β J.RT3-T3.5 Jurkat细胞，或通过用逆转录病毒载体感染表达为正常细胞毒性T淋巴细胞（CTL）系。 B.为了确定这些具有克隆扩增的TCR的转导或转染的T细胞是否识别推定的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.为了进一步确定整个AAA T细胞浸润的功能并确保获得功能响应性T细胞克隆，我们将通过有限稀释法开发对推定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。 a.为了充分鉴定这些T细胞克隆。 B.将这些T细胞克隆使用的TCR序列与来自相同患者的新鲜（未在培养物中扩增）AAA浸润性T细胞的TCR序列进行比较，以确定这些新鲜浸润性T细胞的克隆群体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病变中趋化因子和Th 1和Th 2细胞因子（转录本和蛋白质）的产生，并鉴定产生这些分子的单个细胞类型。 将浸润程度、浸润T细胞的活化阶段、T细胞寡克隆群的存在以及AAA病变中趋化因子和细胞因子的产生与该交互式RO 1计划的第二个RO 1研究资助申请的结果相关联，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。