IN VIVO STUDIES OF SALIVA ANTIMICROBIAL PROTEINS

唾液蛋白可能通过以下机制影响牙菌斑 细胞被杀死，细菌聚集，并附着在膜上。在……里面 体外研究已经确定了参与每个过程的唾液蛋白。 然而，很难证明特定的蛋白质是如何影响 活体内斑块成分。唾液蛋白不同的人 通常会比较不同的水平。这种比较中的一个问题是， 唾液蛋白质形成的复合体的功能可能与 它们的组件。许多蛋白质也显示出多余的功能。那些 问题使我们很难分离唾液蛋白在 人类受试者。这个项目将采用一种新的方法来实现这一点 有问题。唾液功能差异很大的人将是 以检验它们在唾液和唾液方面不同的假设 斑块成分。这一目标将通过以下方式实现 具体目标：1.)开发细胞定量的微孔板方法- 杀戮、聚合和坚持。2.)将这些方法应用于筛选 170名牙科学生未受刺激的全唾液。多变量 分析将被用来对相反的受试者进行细胞杀灭， 聚合和坚持。3.)通过图像分析比较分组 天然的和还原的凝胶来测试冗余和 络合作用将使不同成分的唾液发挥作用 类似的效果。生物素化的蛋白质覆盖物将用于比较 参与聚集和粘连的蛋白质和复合体。结果 来自1-D凝胶将指导使用2-D凝胶或 特异蛋白检测。4.)测试唾液蛋白的假设 在体内发挥选择性作用。牙菌斑会在牙釉质上堆积 放置在唾液导管附近的芯片。一个任意的- 将采用引物聚合酶链式反应方法鉴定链球菌 认为本土植物区系在不同群体之间存在差异的假设 唾液的功能不同。唾液涂布的芯片将被 识别淀粉酶-SIgA复合体的抗独特型抗体，以检测 认为冗余将抑制殖民的假说 通过具有相应受体的细胞。细菌包裹的芯片将被 用受体的原始抗体处理，以测试 淀粉酶与缺乏淀粉酶受体的细胞结合的假说是 由淀粉酶-SIgA复合体介导。建议的工作直接涉及 唾液是斑块的生态决定因素的假说，以及 有可能证实体外研究表明这种 关系是存在的。通过关注冗余性和复杂性，这 该项目还解决了单个蛋白质是否 在口腔中发挥独特而特定的效果。

Salivary proteins may influence dental plaque through mechanisms such as cell killing, bacterial aggregation, and adherence to pellicle. In vitro studies have identified saliva proteins involved in each process. However, it has been difficult to show how particular proteins affect plaque composition in vivo. Persons with different saliva protein levels typically are compared. One issue in such comparisons is that saliva proteins form complexes which may function differently than their components. Many proteins also show redundant functions. Those problems make it difficult to isolate the role of salivary proteins in human subjects. This project will employ a novel approach to that problem. Persons who differ greatly for saliva functions will be identified, to test the hypothesis that they differ for saliva and plaque composition. That goal will be addressed through the following specific aims: 1.) Develop microplate methods for quantifying cell- killing, aggregation, and adherence. 2.) Apply those methods to screen unstimulated whole saliva from 170 dental students. Multivariate analysis will be used to group subjects opposite for cell-killing, aggregation, and adherence. 3.) Compare groups by image analysis of native and reducing gels to test the hypothesis that redundancy and complexing will allow salivas with different compositions to exert similar effects. Biotinylated protein overlays will be used to compare proteins and complexes involved in aggregation and adherence. Results from 1-D gels will direct further comparisons using 2-D gels or specific protein assays. 4.) Test hypotheses that saliva proteins exert selective effects in vivo. Plaque will be accumulated on enamel chips placed at sites adjacent to salivary ducts. An arbitrarily- primed PCR method for streptococcal identification will be used to test the hypothesis that indigenous floras differ between groups whose salivas function differently. Saliva coated chips will be treated with anti-idiotype antibodies recognizing an amylase-sIgA complex, to test the hypothesis that redundancy will present inhibition of colonization by cells with a corresponding receptor. Bacteria-coated chips will be treated with the original antibody to the receptor, to test the hypothesis that amylase binding to cells lacking amylase receptors is mediated by amylase-sIgA complexes. Proposed work directly addresses the hypothesis that saliva is an ecological determinant of plaque, and has the potential to validate in vitro studies which suggest that such a relationship exists. By focusing on redundancy and complexing, this project also addresses the question of whether individual proteins exert unique and specific effects in the mouth.