Basal-Lateral/Endomembrane Traffic in Lacrimal Acini

描述(申请人提供)：原发性泪腺发育不全和干燥综合征 自身免疫性泪腺炎在女性中比男性更常见，而且 荷尔蒙因素的组合，即生物可利用的睾丸激素不足 和过多的催乳素，可能会导致这两种疾病。泪腺 屈光手术后也可能发生眼球功能不全。肖格伦(氏)自身免疫 靶向M3 M受体，一种细胞表面蛋白，以及各种 胞内蛋白质。对两国之间的交通有了新的理解 基侧质膜和内膜室提示 免疫系统可能会对熟悉的蛋白质失去耐受性，或被激活 当：(A)通信量的变化以下列方式增加速率时， 哪些腺泡细胞在其BLM处表达细胞内自身抗原或分泌 (B)腺泡细胞开始表达MHC-II类 分子并将自身抗原肽直接呈递给CD4T细胞；或(C) 腺泡细胞分泌到间质的免疫调节因子谱 变成免疫刺激而不是免疫抑制。活体实验 改变BLM/内膜流量的扰动包括：慢性和 超极量分泌刺激，可能在体内出现为泪液 腺眼表面伺服机制对泪道功能障碍的补偿作用 雄激素不足或催乳素过多；用表皮刺激 生长因子(EGF)，可在体内诱导对角膜的反应 创伤；以及在催乳素存在的情况下进行培养，这可能会模拟高 催乳素状态。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 腺泡细胞用表皮生长因子(EGF)刺激。 2.绘制M3胆碱能受体的流量图；检验腺泡的假设 细胞向间质分泌M3蛋白水解物；腺泡细胞 通过MHC II类分子将M3多肽递送到CD4T细胞； 卡巴胆碱的刺激可能会降低M3受体的周转 减少M3相关抗原刺激。 3.测试含有过量催乳素的环境会改变的假设 无胆碱能刺激和有胆碱能刺激时的蛋白质分选 过量的催乳素增加M3受体的周转，潜在地增加 M3相关抗原刺激。 4.定位内源性表达的IL-2并转导的流量 抗炎细胞因子，如IL-10。测试白介素10的假设 改变内源性蛋白质的运输；卡巴胆碱的慢性刺激 在没有雄激素的情况下，IL-2和IL-10通过心尖部的交通增加 分泌途径；以及在存在的情况下慢性卡巴胆碱刺激 雄激素通过BLM增加分泌。

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.