DEVELOPMENT OF A CHECKERBOARD IMMUNOBLOT

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 有几条证据表明，免疫机制是牙周病组织破坏的特征。 龈沟液（GCF）可以提供一种非侵入性的诊断手段，以衡量炎症介质释放过程中的疾病进展。 在GCF中检测到几种免疫介质，其水平与疾病状态相关。 我们建议使用棋盘免疫印迹（CBIB）技术来量化的GCF在大量的样品中的多个组件。 在我们的实验室中已经应用了类似的方法来表征与牙周感染相关的微生物群。 这项技术将使我们能够用微生物数据来表征GCF组件。 没有这样的高通量技术已可用于细胞因子或其他标志物的宿主状态在牙周网站。 初步数据表明，将有可能在常规GCF样本中测量大量细胞因子。 因此，应该可以在个体内逐部位量化细菌挑战的性质和宿主反应的性质。 一些炎症介质的水平可以影响其他炎症介质的合成和分泌。 同时研究几种分子的能力将允许推断这些介质调节免疫应答和组织破坏的机制。 这种方法将填补空白，我们的知识，关于炎症介质的交叉调节和这些介质的水平，龈下牙周病原体的关系。

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.