T LYMPHOCYTE DYSFUNCTION IN LUPUS ERYTHEMATOSUS

红斑狼疮 T 淋巴细胞功能障碍

• 批准号: 2079548
• 负责人: GARY M KAMMER
• 金额: $ 16.48万
• 依托单位: WAKE FOREST UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-09-01 至 1997-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2079548
• 关键词: T lymphocyte; disease /disorder proneness /risk; enzyme activity; enzyme deficiency; family genetics; human subject; isozymes; molecular pathology; phosphotransferases; protein isoforms; protein kinase A; protein structure function; systemic lupus erythematosus

We have described a disorder of cAMP metabolism in T lymphocytes of subjects with systemic lupus erythematosus (SLE) Our recent identification of defective cAMP -dependent phosphorylation of proteins in intact SLE T lymphocytes suggests a disorder of cAMP -dependent protein kinase (Protein kinase A) The objective of this proposal is to identify and characterize the abnormal protein kinase A function in SLE T lymphocytes by quantifying cAMP -dependent protein kinase activities and the amounts of the enzyme's constituent subunits, the types I (RI) and II (RII) regulatory and catalytic subunits (C subunit). The specific aims of this proposal are (a) to compare protein kinase A activities in the nonparticulate and particulate fractions of T lymphocytes and T lymphocyte subsets from active and inactive SLE subjects with controls. cAMP -dependent protein kinase activity will be quantified by measuring the phosphorylation of histone and Kemptide in nonparticulate and particulate T cell extracts. (b) We will purify partially the cAMP -dependent protein kinases of SLE T lymphocytes by DEAE -cellulose chromatography in order to determine the presence of anomalous RI, RII and/or C subunits. Anomalous RI or RII subunits can exhibit altered cAMP binding while anomalous C subunits can possess reduced binding affinity for ATP. (c) The amounts of protein kinase A subunits will be quantified by ELISA and nitrocellulose immunolabeling to determine whether a deficiency of one or more subunits exists to explain altered protein kinase activities. Differences in the kinase activities and/or amounts of a kinase of a kinase subunit between T cell subsets will be quantified to determine whether a restricted kinase defect exists. Since the T lymphocyte plays an integral role in both cellular and humoral immunity, it is important to discover whether a defective cAMP pathway accounts, in part, for the aberrant immunoregulation in SLE. Thus, our longterm goal is to determine whether defective protein kinase A function results in abnormal suppressor cell activity. The identification of a protein kinase A defect should provide important insights into the pathogenesis of this autoimmune disorder.

我们描述了T细胞cAMP代谢紊乱， 系统性红斑狼疮（SLE）患者的淋巴细胞 我们最近发现的缺陷型cAMP依赖性 完整SLE T淋巴细胞中蛋白质的磷酸化表明 cAMP依赖性蛋白激酶（蛋白激酶A）疾病 本提案的目的是确定和描述 SLE T淋巴细胞蛋白激酶A功能异常 定量cAMP依赖性蛋白激酶活性， 酶的组成亚基，I型（RI）和 II（RII）调节和催化亚基（C亚基）。 具体 该建议的目的是（a）比较蛋白激酶A T的非颗粒和颗粒组分中的活性 淋巴细胞和T淋巴细胞亚群从活动和非活动 SLE受试者与对照。 cAMP依赖性蛋白激酶 活性将通过测量磷酸化 非颗粒和颗粒T细胞中组蛋白和Kemptide 萃取物(b)我们将部分纯化cAMP依赖性蛋白 SLE T淋巴细胞激酶的DEAE-Cellulose层析 为了确定异常RI、RII和/或C的存在， 亚单位。 异常的RI或RII亚基可以表现出cAMP的改变 结合，而异常C亚基可以具有降低的结合 对ATP的亲和力。 (c)蛋白激酶A亚单位的量 将通过ELISA和硝酸纤维素免疫标记进行定量， 确定是否存在一个或多个亚单位的缺陷， 解释蛋白激酶活性的改变 差异 激酶亚基的激酶活性和/或激酶量 将对T细胞亚群之间的差异进行定量，以确定是否存在免疫缺陷。 存在限制性激酶缺陷。 由于T淋巴细胞起着 在细胞免疫和体液免疫中起着不可或缺的作用， 重要的是要发现是否有缺陷的cAMP途径 部分解释了SLE中异常的免疫调节。 因此，我们的长期目标是确定缺陷蛋白质是否 激酶A功能导致抑制细胞活性异常。 蛋白激酶A缺陷的鉴定应提供 对这种自身免疫性疾病的发病机制的重要见解 disorder.