翰斯勒巴東體分離株感染內皮細胞致病機轉之研究; The study of cellular responses between different Bartonella henselae strains

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Title:	翰斯勒巴東體分離株感染內皮細胞致病機轉之研究;The study of cellular responses between different Bartonella henselae strains
Authors:	陳雅柔;Ya-Jou Chen
Contributors:	中國醫藥大學：生物科技學系碩士班
Keywords:	翰斯勒巴東體;內皮細胞;致病機轉;Bartonella henselae;endothelial cells;proteomic analysis;pathogenesis
Date:	2009-06-09
Issue Date:	2009-08-11 10:58:36 (UTC+8)
Abstract:	翰斯勒巴東體 (Bartonella henselae) 是革蘭氏陰性菌，會造成人畜共通傳染病。貓為保菌者 (reservoir)，為引起貓抓熱 (cat-scratch disease) 的病原菌，對免疫不全症候群的病人會造成桿菌性血管瘤。我們將4株B. henselae感染細胞後進行比較，發現此菌經由抑制粒線體內在凋亡路徑，進而促使細胞增生；且不同分離株間促進細胞增生及抑制凋亡的能力不盡相同，Houston-1 (Hous) 分離株的能力最強，其次為JK40與U-4，最弱為JK47。於刺激ROS產生的實驗中，只有JK47的能力較差，而Hous具有最強的細胞侵入能力。發炎反應及細胞黏附能力部分，genotype I 的Hous及JK47較genotype II的U-4及JK40強。在細胞反應的所有實驗中，Hous分離株均具有最強的能力，並於蛋白質體分析中，發現small heat shock protein、acetyl-CoA carboxylase carboxyltransferase subunit alpha、phage related protein及superoxide dismutase [Cu-Zn] precursor等蛋白質，在感染後於不同分離株之RNA表現量與各分離株刺激細胞反應強度具有一致性。此項研究為首次以系統性的方式比較B. henselae分離株所引起的細胞反應與蛋白質圖譜，我們同時發現與B. henselae感染細胞後所引起的細胞反應的相關蛋白質，其極可能是造成致病性的重要因子。 Bartonella henselae is an agent capable of causing a wide variety of disease syndromes and is an emerging pathogen that causes potentially fetal opportunistic infection in patients with acquired immunodeficiency syndrome. The most common B. henselae-associated diseases are cat-scratch disease and bacillary angiomatosis. In this study, the interaction between B. henselae strains and endothelial cells was studied and the candidates in response to the pathogenesis were also determined. We identified that B. henselae infection inhibited the mitochondria intrinsic pathway and the strain stimulated more cell proliferation would suppress more intrinsic apoptotic pathway. Houston-1 (Hous) strain possessed the strongest ability in stimulating cell proliferation and inhibiting infection-induced apoptosis. Further more, Hous, U-4 and JK40 induced more ROS production than JK47 did. In bacterial adhesion abilities, genotype II (U-4 and JK40 strains) showed better adhesion abilities than genotype I (Hous and JK47). However, the invasion ability of Hous was the best among all the strains. Hous and JK47 (genotype I) could also induce more IL-8 (proinflammatory factor) production than U-4 and JK40 (genotype II) did. In proteomic analysis, two-dimensional gel electrophoresis and real-time PCR analysis were used to identify virulence factors in B. henselae. The mRNA levels of small heat shock protein, succinyl-CoA synthetase subunit beta, phage related protein and superoxide dismutase [Cu-Zn] precursor were increased in B. henselae after infection. These proteins might play important roles in pathogenesis of B. henselae.
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