APOPTOSIS IN SJOGRENS SYNDROME

干燥综合征中的细胞凋亡

• 批准号: 2628887
• 负责人: Antony Rosen
• 金额: $ 27.15万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-05-01 至 2002-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2628887
• 关键词: HeLa cells; SDS polyacrylamide gel electrophoresis; Sjogren's syndrome; active sites; apoptosis; autoantigens; autoimmune disorder; biopsy; cell type; clinical research; cytotoxic T lymphocyte; cytotoxicity; dendritic cells; endopeptidases; gastrointestinal epithelium; human subject; immunocytochemistry; recombinant proteins; salivary glands; western blottings

DESCRIPTION (Adapted from investigator's Abstract): Sjogren's syndrome (SS) is a chronic autoimmune disease which results in decreased secretion from salivary and lacrimal glands. Although intense lymphocytic infiltration and epithelial cell destruction are characteristic of the disease, identifying the actual mechanism(s) of salivary epithelial injury in SS remains a major challenge. The broad, long-term objectives of this proposal are to define salivary gland epithelial cell apoptosis in SS, and to address the role of granule-mediated cellular cytotoxicity in this process. The specific aims of the proposal are: (1) to quantify and characterize salivary gland epithelial cell apoptosis in SS, and to demonstrate the novel autoantigen fragments specifically generated by the granule-mediated cytotoxicity pathway in SS salivary glands; (2) to use the human salivary gland cell line (HSG) as a model system to study the consequences of cytolytic granule-induced apoptosis on autoantigen structure and distribution; (3) to identify the sites at which granzyme B cleaves La; and (4) to address immunogenicity of novel La fragments in vitro. Apoptosis in salivary gland biopsies from SS patients will be quantitated using the TUNEL assay, and will be correlated with the lymphocytic infiltrate, both in terms of distribution and magnitude. Vectorial distribution of cytolytic T cell granules will be sought in SS salivary glands. The apoptotic pathways induced by ultraviolet irradiation and cytolytic granule contents in HSG cells and HeLa cells will be compared. The granzyme B cleavage site in La will be defined by radiosequencing or mutational analysis. Recombinant proteins corresponding to the intact protein and the granzyme B cleavage fragments will be generated, and their immunogenicity will be addressed using an in vitro immunogenicity assay. These studies will yield an enhanced understanding of the relevance of granule-mediated apoptosis in the pathogenesis of SS and other autoimmune diseases.

描述（改编自调查员的摘要）：Sjogren综合征（SS） 是一种慢性自身免疫性疾病，导致从 唾液和泪腺。 虽然强烈的淋巴细胞浸润和 上皮细胞破坏是该疾病的特征，确定 SS中唾液上皮损伤的实际机制仍然是主要的 挑战。 该提议的广泛，长期目标是定义 SS中的唾液腺上皮细胞凋亡，并解决 在此过程中，颗粒介导的细胞毒性。 具体目标 该提案的内容是：（1）量化和表征唾液腺 SS中的上皮细胞凋亡，并演示新型自身抗原 由颗粒介导的细胞毒性专门产生的片段 SS唾液腺中的途径； （2）使用人类唾液腺细胞系 （HSG）作为研究细胞溶解后果的模型系统 颗粒引起的自身抗原结构和分布的凋亡； （3）到 识别颗粒状B裂的位置； （4）解决 新型LA碎片体外的免疫原性。 唾液腺凋亡 SS患者的活检将使用TUNEL分析进行定量，并且 都将与淋巴细胞浸润相关 分布和大小。 细胞溶解T细胞的矢量分布 SS唾液腺将寻求颗粒。 凋亡途径 HSG中的紫外线照射和细胞溶解颗粒含量诱导 将比较细胞和HeLa细胞。 LA中的Granzyme B裂解部位 将通过放射测序或突变分析来定义。 重组 对应于完整蛋白质和颗粒状B裂解的蛋白质 将产生碎片，并将其免疫原性解决 使用体外免疫原性测定法。 这些研究将产生 对颗粒介导的凋亡相关性的了解增强了 SS和其他自身免疫性疾病的发病机理。