2017-08-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/689454摘要：非結核分枝桿菌引起之感染於全球有不斷增加之趨勢。其中，皮膚及軟組織為肺部外最常見之感染部位，其臨床及組織病理學上之表現隨著宿主免疫力及菌種的不同會呈現個別差異，因此缺乏典型之臨床診斷依據。傳統實驗室方法診斷率很低，再加上治療須使用多種藥物組合、療程冗長且藥物副作用大，不同菌種（甚至亞種）對藥物之感受性亦不盡相同，某些菌種又易於造成群突發，所以這類感染已成為臨床醫療之棘手課題。然而，截至目前為止，學界對相關議題的探討卻甚為匱乏。緣此，為解決這些臨床診斷及治療上之困境，我們擬橫向結合臨床、流行病學、病理學及免疫學各種面向，縱以病人檢體、細胞株培養及動物實驗等模式，詳細去釐清疾病機轉及驗證假說。首先，我們將收集過去7 年已確診為分枝桿菌皮膚及軟組織感染病人之臨床菌株，以分子生物學方法作菌種及亞種鑑定(尤其是M. abscessus及Ｍ. massiliense，後者近年常見於群突發感染），分析不同菌種及亞種之感染於臨床、病理學及藥敏試驗結果之差異，期能找出有助於臨床醫師提高警覺及初步區別感染原因之依據。其次，為找出更快捷、更敏感及更準確之診斷方法，我們前瞻性地同步收集臨床上難治之皮膚及軟組織感染病人之切片，以聚合酶鏈鎖反應(PCR)偵測皮膚組織檢體內是否具有分枝桿菌，並輔以定序來鑑別菌種（與亞種）。我們會同時分析病人血中是否會產生一種對抗分枝桿菌醣類胺基酸脂質鍵結物（glycopeptolipid)之免疫球蛋白，再比較這些分子生物學及血清學診斷方法是否具有優於傳統培養及病理診斷之效能(performance)。第三，我們會利用細胞培養，探討不同菌種感染角質細胞株之能速度、細胞內存活能力、透發活性氧化物質、細胞激素、抗微生物胜肽等之情形。最後，我們將根據臨床及細胞實驗分析之結果，建立動物模型來驗證組織免疫學假說。藉由整合這些臨床及基礎之研究結果，我們期能闡明不同分枝桿菌與宿主間之交互影響外，亦能建立未來開發更快速與更精準診斷，以及更有效治療策略之轉譯研究平台。<br> Abstract: Non-tuberculous mycobacterial (NTM) infections are increasingly reported worldwide. Skin andsoft tissue infections (SSTIs) are the most common extrapulmonary diseases caused by NTM.Diagnosis of mycobacterial SSTI can be challenging due to diverse clinical presentation, low yieldfrom cultured specimens, and nonspecific histopathology on tissue biopsy. Despite aggressive medicaltreatment that necessitates prolonged combination therapy with frequent drug toxicities, responses maybe suboptimal with poor outcomes. The precise differentiation of the etiologic organisms is crucial forthe clinicians since the treatment response and susceptibility to drugs differ among subspecies. Basedon our previous work and preliminary clinical and in vitro study results, we also found that the tissuetropism pathogenic potential of NTM differs substantially by species and even subspecies. Given theunmet medical needs of rapid diagnosis, speciation and better treatment strategies, we will conduct amultifaceted study, utilizing clinical investigations, in vitro and in vivo models, to better understand thedistinctive clinical features, histopathological characteristics, and the immunopathogenesis triggered bydifferent strains of NTM in the cutaneous microenvironment. First, we will seek to identify usefulclinical clues to assist clinicians in distinguishing SSTIs caused by different NTM strains. Storedclinical strains will be collected and further speciated by molecular methods. We will then compare thedemographics, clinical characteristics, histopathological features and susceptibility patterns of variousstrains of NTM isolated from the infected sites, with special focus on M. abscessus versus M.massiliense (an emerging NTM causing nosocomial outbreaks). Second, in order to determine whetherPCR can expedite laboratory diagnosis and speciation accurately, we will prospectively collect biopsyspecimens from patients with protracted SSTIs for PCR and DNA sequence analyses. Serum will alsobe obtained to detect the level of anti-glycopeptolipid immunoglobulins. Results of the molecularassays will be compared with the performance of conventional culture and histopathologicalexamination. Third, we will investigate the infectivity, differences in the productions of reactiveoxygen species (ROS) and proinflammatory cytokines in human keratinocytes infected with variousspecies of NTM. Finally, we aim to establish a mouse model to decipher host-microbe interactions invivo. The results from these integrated, in-depth investigations are anticipated to pave ways towardunderstanding the pathogenesis, development of more timely and accurate diagnosis, as well as moreeffective treatment strategies for NTM infections.分枝桿菌皮膚及軟組織感染免疫組織病理學nontuberculous mycobacterial infectionNTMskin and soft tissue infectionsimmune responsehistopathological