INTERLEUKIN-2 PRODUCTION IN SJOGREN'S SYNDROME

摘要原发性干燥综合征是一种慢性自身免疫性疾病。 其特征是1)血清自身抗体，2)CD5+B细胞数量增加 细胞，3)唾液、泪液和其他外分泌的淋巴细胞浸润 有全身性淋巴组织增殖倾向的腺体 以恶性淋巴瘤的形式终止。这样做的长期目标是 应用是为了获得知识，从而1)更好地理解 SS免疫失调的机制：2) 可能开发新的治疗方式(如多胺 抑制剂)治疗这种疾病。外周血单个核细胞(PBMNC) SS患者产生的IL-2量减少 用PHA或抗CD3刺激后。还有一个跨膜的 暴露在PMA和离子霉素下的SS T细胞信号缺陷。 由于多胺的氧化产物在正常情况下会减少IL-2的产生 和类风湿关节炎(RA)T细胞，可能有类似的机制在运作 和初级SS。具体目标和方法如下： 1.测量多胺水平(用高效液相色谱法)、白介素2产量(用生物测定法)和 多胺抑制剂(如DEMO)对SS细胞产生IL-2的影响 PBMNC。 2.通过狭缝杂交和Northern杂交检测SS PBMNC中IL-2的mRNA水平 确定可能的转录前或转录后缺陷(水解酶 离子交换层析测定肌醇磷脂在细胞内的含量 Indo-1和FACS分析的钙离子，二酰甘油依赖的激活， IL-2蛋白的生物合成和分泌(用ELISA法)。 3.尝试通过多胺抑制剂纠正SS中可能存在的缺陷，或 使用多胺加多胺氧化酶在正常的PBMNC中产生缺陷。

Primary Sjogren's syndrome (SS) is a chronic autoimmune disease characterized by 1) serum autoantibodies, 2) increased numbers of CD5+ B cells, 3) lymphocytic infiltration of salivary, lacrimal and other exocrine glands with a tendency to generalized lymphoproliferation that can terminate as a malignant lymphoma. The long-term objective of this application is to acquire knowledge leading to 1) a better understanding of the mechanisms responsible for the immune dysregulation in SS, and 2) possible development of new therapeutic modalities (such as polyamine inhibitors) for this disease. Peripheral blood mononuclear cells (PBMNC) from SS patients produce decreased amounts of interleukin-2 (IL-2) following stimulation with PHA or anti-CD3. There is also a transmembrane signalling defect in SS T cells revealed by exposure to PMA and ionomycin. Since oxidation products of polyamines diminish IL-2 production by normal and rheumatoid arthritis (RA) T cells, a similar mechanism may be operating and primary SS. The specific aims and methodologies are the following: 1. Measure polyamine levels (by HPLC), IL2 production (by bioassay) and the effect of polyamine inhibitors (like DEMO) on IL2 production in SS PBMNC. 2. Measure IL2 mRNA levels in SS PBMNC (by slot and Northern blots) and identify possible pre- or post-transcriptional defects (hydrolysis of phosphoinositides by ion exchange chromatography, rise in intracellular calcium by Indo-1 and FACS analysis, diacylglycerol-dependent activation, IL2 protein biosynthesis and secretion by ELISA). 3. Attempt to correct a possible defect in SS by polyamine inhibitors, or create a defect in normal PBMNC using polyamines plus polyamine oxidase.