Identification, recombinant expression, immuno localization in macrophages, and T-cell responsiveness of the major extracellular proteins of Francisella tularensis
Identification, recombinant expression, immuno localization in macrophages, and T-cell responsiveness of the major extracellular proteins of Francisella tularensis
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DOI：
10.1128/iai.00257-06
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发表时间：
2006-07-01
影响因子：
3.1
通讯作者：
Clemens, Daniel L.
中科院分区：
文献类型：
--
作者：
Lee, Bai-Yu;Horwitz, Marcus A.;Clemens, Daniel L.
需要一种比以前开发的减毒活疫苗更安全和更有效的疫苗来对抗土拉热弗朗西丝菌，一种高度传染性的细菌病原体。为了寻找包含在新疫苗中的潜在候选者，我们表征了存在于最近的临床分离株和F. tularensis使用蛋白质组学方法。我们确定了共12种蛋白质，其中，过氧化氢酶-过氧化物酶是更丰富的培养滤液中的毒性临床分离株，而细菌铁蛋白是更丰富的活疫苗株的培养滤液。链球菌溶血素0处理感染的人巨噬细胞表明，过氧化氢酶-过氧化物酶和热休克蛋白GroEL是由活跃生长的F。土拉热。用F.通过脾淋巴细胞增殖和γ-干扰素产生来测量，土拉热菌对过氧化氢酶-过氧化物酶、热休克蛋白GroEL和细菌铁蛋白产生了显著的细胞介导的免疫应答。最后，我们在大肠杆菌中以可溶性形式高水平表达了有希望的疫苗候选物的主要培养滤液蛋白，以促进其免疫生物学和在疫苗中的潜在作用的研究。
A safer and more effective vaccine than the previously developed live attenuated vaccine is needed for combating Francisella tularensis, a highly infectious bacterial pathogen. To search for potential candidates for inclusion in a new vaccine, we characterized the proteins present in the culture filtrates of a virulent recent clinical isolate and the attenuated live vaccine strain of F. tularensis using a proteomic approach. We identified a total of 12 proteins; among these, catalase-peroxidase was much more abundant in the culture filtrate of the virulent clinical isolate, whereas bacterioferritin was more abundant in the culture filtrate of the live vaccine strain. Streptolysin 0 treatment of infected human macrophages indicated that catalase-peroxidase and the heat shock protein GroEL are released intracellullarly by actively growing F. tularensis. Mice immunized with F. tularensis developed significant cell-mediated immune responses to catalase-peroxidase, the heat shock protein GroEL, and bacterioferritin as measured by splenic lymphocyte proliferation and gamma interferon production. Finally, we expressed the major culture filtrate proteins that are promising vaccine candidates in Escherichia coli at high levels in soluble form to facilitate study of their immunobiology and potential role in vaccines.