Burkholderi pseudomallei 屬兼性胞內寄生菌， 可藉由macrophages 之 internalization 時逃脫phgagosome 而侵入細胞內。並可誘導感染細胞核融合，產生獨特的多何巨大細胞(MNGC) ; 和誘導細胞生caspase-1-dependent apoptosis 。但迄今對Burkholderi pseudomallei 如何逃脫macrophages 之ROI (reactive oxygen intermediates) 與RNI (reactive nitrogen intermediates) 的 oxidative killing mechanism，以及如何誘導macrophage 之apoptosis 的發生，仍不清楚。本計劃旨在分析Burkholderi pseudomallei (WT, fliC-, wbiI- or rpoS-)感染 macrophage-like cell lines 後相關signaling pathway (如Toll-like pathway (TLR pathway) 和apoptotic pathway; 這些pathway 已被證實與菌種抗吞噬作用有關)，然而，無任何文獻探討這些pathway 上，中，下游蛋白質是否改變? 及感染細胞內mi RNA 改變，是否影響macrophage 吞噬能力? 這些研究將以 high-throughput 研究方法如PCR array and proteome，以[全面性] 釐清菌體的致病機制。並發現感染細胞一些獨特基因或蛋白質可作為診斷之生物指標。Burkholderi pseudomallei can be resistant to phagocytosis, cause the occurrence of caspase-1-dependent apoptosis in macrophage, and induce the formation of multinuclear giant cell (MNGC). It is uncertain how B. pseudomallei escape from the attacking of ROI- (reactive oxygen intermediates) or RNI- (reactive nitrogen mediated oxidative killing mechanism that produced by macrophages. No research focused on the detailed mechanism of apoptosis or toll-like pathway, an important pathway for reorganization of pathogens. In addition, the involvement of miRNA in regulation of phogocytosis was suggested herein. In this project, the downstream and upstream of toll-like pathway-, apoptotic protein-related gene cascade is studied as comparative studies using Burkholderi pseudomallei mutants (fliC-, wbiI- or rpoS-) and counterparts. The use of high-throughput techniques such as PCR array or proteome can enable us to unravel many of differential genes or proteins simultaneously. Thus, the aim of this study is to delineate the mechanisms of bacterial resistance to phagocytosis, and find out the differential genes or protein in macrophages before and after infection. These genes or proteins could be a potentially diagnostic biomarker.研究期間：10108 ~ 10207
