SUBEPITHELIAL ANTIGENS

上皮下抗原

• 批准号: 2542941
• 负责人: ROBERT E BURGESON
• 金额: $ 35.72万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-09-23 至 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2542941
• 关键词: artificial skin; basement membrane; biological models; cell adhesion; cell adhesion molecules; epidermolysis bullosa; epithelium; human tissue; intercellular connection; keratinocyte; laboratory mouse; laboratory rabbit; laminin; protein sequence; protein structure function; skin

The anchoring complex is an ultrastructural feature of the dermal-epidermal basement membrane that includes the hemidesmosomes, anchoring filaments and anchoring fibrils. These structures are believed to work as a unit to stabilize epithelial attachment to the underlying basement membrane. Failure of components of the anchoring complex results in spontaneous blistering characteristic of the inherited bullous diseases. We have recently identified two laminin variants that are components of the anchoring filaments. We have termed these kalinin and K-laminin. We know that kalinin is absent from the basement membrane zone of patients with lethal epidermolysis bullosa. Kalinin is a cell adhesion macromolecule specific for certain types of epithelial cells including keratinocytes. For these cells, kalinin substitutes for laminin as the favored growth substrate. Monoclonal antibodies made to kalinin cause cell detachment and de-epithelialization of whole skin in vitro. This application is focused upon continued characterization of the structure and function of kalinin and K-laminin. We propose to isolate kalinin and K-laminin for protein sequence determinations and for preparing protein fragments; to complete the deduced amino acid sequences of these chains and to express selected subdomains in vitro; and to generate domain specific monoclonal antibodies. These reagents will be used to evaluate the binding function of kalinin and K-laminin subdomains. We will interfere with these functions in an in vitro skin equivalent model. These probes will also be used to evaluate mutations in the kalinin and K-laminin chains relative to the pathogenesis of junctional epidermolysis bullosa.

锚定复合体是一种超微结构特征 真皮-表皮基底膜，包括半桥粒， 锚定细丝和锚定纤维。这些结构被认为 作为一个单位工作，以稳定上皮对底层的附着 基底膜。锚固复合体的部件发生故障 导致遗传性大疱病的自发性水泡特征 疾病。我们最近发现了两种层粘连蛋白变体，它们是 锚定细丝的组成。我们把它们命名为加里宁和 K-层粘连蛋白。我们知道加里宁不在基底膜上 致死性大疱性表皮松解症患者的区域。加里宁是一个细胞 某些类型的上皮细胞所特有的黏附大分子 包括角质形成细胞。对于这些细胞，加里宁取代了 层粘连蛋白是最受青睐的生长底物。抗人免疫球蛋白单抗 加里宁引起大鼠全皮肤细胞脱落和脱上皮化 体外培养。本申请的重点是继续表征 加里宁和K-层粘连蛋白的结构和功能。我们建议 分离的Kalinin和K-laminin用于蛋白质序列测定和 制备蛋白质片段，完成推导的氨基酸序列 并在体外表达选定的亚区； 产生区域特异性的单抗。这些试剂将是 用来评价卡尔宁和K-层粘连蛋白的结合功能 子域。我们将在体外皮肤中干扰这些功能 等效模型。这些探针也将被用来评估突变 在Kalinin和K-laminin链中与糖尿病的发病机制有关 交界性大疱性表皮松解症。