DEVELOPMENT OF A CHECKERBOARD IMMUNOBLOT

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 一些证据表明，免疫机制是牙周病特有的组织破坏的原因。牙周沟液(GCF)可以提供一种非侵入性的诊断手段来检测疾病进展过程中所释放的炎性介质。已在GCF中检测到几种免疫介质，它们的水平与疾病状态有关。 我们建议使用棋盘免疫印迹(CBIB)技术来定量大量样本中GCF的多种成分。一种类似的方法已经在我们的实验室中用于表征与牙周感染相关的微生物区系。这项技术将使我们能够用微生物数据来表征GCF的组成部分。目前还没有如此高通量的技术可用于牙周部位的细胞因子或其他宿主状态的标志物。 初步数据表明，有可能在常规的GCF样本中测量大量的细胞因子。因此，应该有可能在个体内逐个部位的基础上量化细菌挑战的性质和宿主反应的性质。某些炎症介质的水平会影响其他炎症介质的合成和分泌。同时研究几个分子的能力将允许对这些介体调节免疫反应和组织破坏的机制进行推断。这种方法将填补我们关于炎症介质的交叉调节以及这些介质的水平与牙周下病原体之间的关系的知识空白。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Several lines of evidence demonstrate that immune mechanisms are responsible for the tissue destruction characteristic of periodontal disease. Gingival crevicular fluid (GCF) could provide a non-invasive diagnostic means to measure the inflammatory mediators released during disease progression. Several immune mediators have been detected in GCF, and their levels have been associated with disease status. We propose to use the checkerboard immunoblotting (CBIB) technique to quantify multiple components of the GCF in a large number of samples. A similar approach has been applied in our laboratory for the characterization of the microbiota associated with periodontal infection. This technology would allow us to characterize GCF components with microbiological data. No such high throughput technique has been available for cytokines or other markers of host status at periodontal sites. Preliminary data indicate that it will be possible to measure significant numbers of cytokines in routine GCF samples. Therefore, it should be possible to quantify the nature of the bacterial challenge and the nature of the host response on a site-by-site basis within the individual. The levels of some inflammatory mediators can influence the synthesis and secretion of others. The ability to investigate several molecules at once would allow inferences on the mechanisms of modulation of immune response and tissue destruction by these mediators. This approach will fill a gap in our knowledge regarding the cross-regulation of inflammatory mediators and the relationship of the levels of these mediators to subgingival periodontal pathogens.