IN VIVO STUDIES OF SALIVA ANTIMICROBIAL PROTEINS

唾液蛋白可能通过以下机制影响牙菌斑， 如细胞杀伤、细菌聚集和粘附于表膜。 在 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如何影响 体内的斑块组成。 不同唾液蛋白的人 通常比较水平。 这种比较的一个问题是， 唾液蛋白形成复合物，其功能可能不同于 其组成部分。 许多蛋白质也显示出冗余功能。 那些 这些问题使得很难分离唾液蛋白在 人类实验对象 该项目将采用一种新颖的方法， 问题. 唾液功能差异很大的人， 鉴定，以检验他们对唾液不同的假设， 菌斑组成。 这一目标将通过以下方式实现： 具体目标：1.）开发定量细胞的微孔板方法- 杀伤、聚集和粘附。2.）的情况。将这些方法应用于屏幕 来自170名牙科学生的无刺激全唾液。 多元 分析将用于对细胞杀伤相反的受试者进行分组， 聚集和粘附。3.）第三章通过图像分析比较各组 天然和还原凝胶，以测试冗余和 复合将允许具有不同成分的唾液发挥 类似的效果。 将使用生物素化蛋白覆盖图进行比较 参与聚集和粘附的蛋白质和复合物。 结果 将指导使用二维凝胶进行进一步比较， 特异性蛋白质测定。 4.）的情况。测试唾液蛋白质 在体内发挥选择性作用。 牙菌斑会在牙釉质上积累 在唾液腺导管附近放置芯片。 任意- 将使用引物PCR方法进行链球菌鉴定， 土著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。 唾液涂层芯片将用 识别淀粉酶-sIgA复合物的抗独特型抗体，以检测 冗余将抑制定殖的假设 由具有相应受体的细胞产生。 涂有细菌的芯片将 用受体的原始抗体处理，以测试 假设淀粉酶与缺乏淀粉酶受体的细胞结合， 由淀粉酶-sIgA复合物介导。 拟议工作直接涉及 假设唾液是牙菌斑的生态决定因素， 具有验证体外研究的潜力，这些研究表明， 关系是存在的 通过关注冗余和复杂性， 该项目还解决了单个蛋白质是否 在口腔中发挥独特而独特的作用。

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.