Basal-Lateral/Endomembrane Traffic in Lacrimal Acini

泪腺腺泡的基底外侧/内膜交通

• 批准号: 6331099
• 负责人: AUSTIN K MIRCHEFF
• 金额: $ 51.6万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-04-01 至 2004-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6331099
• 关键词: Golgi apparatus; MHC class II antigen; Sjogren's syndrome; acinar cell; antigen presentation; apical membrane; autoantigens; autoimmune disorder; autoimmunity; basolateral membrane; carbachol; cholinergic receptors; confocal scanning microscopy; cytokine; density gradient ultracentrifugation; electron microscopy; epidermal growth factor; immunocytochemistry; immunoprecipitation; intracellular transport; keratoconjunctivitis sicca; laboratory rabbit; lacrimal apparatus; membrane permeability; protein transport; western blottings

DESCRIPTION (provided by applicant): Primary Lacrimal Deficiency and Sjogren's autoimmune dacryoadenitis are more common in women than men, and the same constellation of hormonal factors, i.e., insufficient bioavailable testosterone and excessive prolactin, may contribute to both pathologies. Lacrimal insufficiency also may occur after refractive surgery. Sjogren's autoimmunity targets the M3 muscarinic receptor, a cell surface protein, and various intracellular proteins. An emerging understanding of traffic between the basal-lateral plasma membranes and endomembrane compartments suggests the immune system may lose its tolerance for familiar proteins, or become activated against cryptic proteins, when: (a) changes in traffic increase the rates at which acinar cells express intracellular autoantigens at their blm or secrete them to the interstitium; (b) acinar cells begin to express MHC Class II molecules and present autoantigen peptides directly to CD4 T cells; or (c) the spectrum of immunomodulatory factors acinar cells secrete to the interstitium becomes immunostimulatory instead of immunosuppressive. In vivo experimental perturbations that alter blm / endomembrane traffic include: chronic and supramaximal secretory stimulation, which might arise in vivo as the lacrimal gland - ocular surface servomechanism compensates for lacrimal dysfunction due to androgen insufficiency or prolactin excess; stimulation with epidermal growth factor (EGF), which may be elicited in vivo in response to corneal trauma; and culture in the presence of prolactin, which may mimic high prolactin states. The specific aims of the proposed work are to: 1. Identify membrane protein sorting mechanisms that are altered when lacrimal acinar cells are stimulated with (EGF). 2. Map the traffic of M3 cholinergic receptors; test the hypotheses that acinar cells secrete M3 proteolytic fragments to the interstitium; that acinar cells present M3 peptides to CD4 T cells via MHC Class II molecules; and that chronic stimulation with carbachol decreases M3 receptor turnover, potentially decreasing M3-related antigenic stimulation. 3. Test the hypotheses that a milieu containing excessive prolactin alters protein sorting in the absence and presence of cholinergic stimulation and that excessive prolactin increases turnover of M3 receptors, potentially increasing M3-related antigenic stimulation. 4. Map the traffic of endogenously expressed IL-2 and transduced anti-inflammatory cytokines, such as IL-10. Test the hypotheses that IL-10 alters traffic of endogenous proteins; that chronic stimulation with carbachol in the absence of androgens increases traffic of IL-2 and IL-10 via the apical secretory pathway; and that chronic carbachol stimulation in the presence of androgen increases secretion via the blm.

描述（由申请人提供）：原发性泪液缺乏症和干燥综合征 自身免疫性泪腺炎在女性中比男性更常见， 激素因素的星座，即，生物可利用睾酮不足 和过多的催乳素，可能会导致这两种病理。泪 屈光手术后也可能发生不全。干燥自身免疫 靶向M3毒蕈碱受体，一种细胞表面蛋白， 细胞内蛋白质一个新兴的理解之间的交通 基底侧质膜和内膜室表明， 免疫系统可能会失去对熟悉蛋白质的耐受性，或被激活 对隐蔽蛋白，当：（a）交通的变化增加率， 腺泡细胞在其BLM表达细胞内自身抗原或分泌 （B）腺泡细胞开始表达MHC II类 分子并直接将自身抗原肽呈递给CD 4 T细胞;或（c）所述抗原肽是抗原肽。 免疫调节因子谱腺泡细胞分泌到胸腺 变得免疫刺激而不是免疫抑制。体内实验 改变BLM /内膜运输的扰动包括：慢性和 超最大分泌刺激，这可能会出现在体内作为泪腺 腺体-眼表伺服机构补偿泪液功能障碍 雄激素不足或催乳素过量;表皮刺激 生长因子（EGF），其可以在体内响应于角膜刺激而被诱发。 创伤;以及在催乳素存在下培养，这可能模拟高 prolactin说。拟议工作的具体目标是： 1.确定泪液分泌时改变的膜蛋白分选机制 腺泡细胞用EGF刺激。 2.绘制M3胆碱能受体的交通图;检验腺泡 细胞分泌M3蛋白水解片段到腺泡;腺泡细胞 通过MHC II类分子将M3肽呈递给CD 4 T细胞; 氨甲酰胆碱的刺激降低了M3受体的转换，可能 减少M3相关的抗原刺激。 3.检验含有过量催乳素的环境改变 在存在和不存在胆碱能刺激的情况下进行蛋白分选， 过量的催乳素增加M3受体的周转， M3相关抗原刺激。 4.绘制内源性表达的IL-2和转导的IL-2的运输图。 抗炎细胞因子，如IL-10。检验IL-10 改变了内源性蛋白质的运输， 在缺乏雄激素的情况下，增加IL-2和IL-10通过顶侧的运输， 分泌途径;以及在存在下慢性卡巴胆碱刺激 雄激素通过BLM增加分泌。