Identification of immunogenic and serum binding proteins of Staphylococcus epidermidis
Identification of immunogenic and serum binding proteins of Staphylococcus epidermidis
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DOI：
10.1128/iai.73.10.6591-6600.2005
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发表时间：
2005-10-01
影响因子：
3.1
通讯作者：
Baker, SM
中科院分区：
文献类型：
--
作者：
Sellman, BR;Howell, AP;Baker, SM
表皮葡萄球菌是人类皮肤的寄生虫，也是医院内血流感染的主要原因。关于S的信息有限。在转移到血流中时表达的表皮蛋白或参与宿主-病原体相互作用的那些。S.通过二维（2D）凝胶电泳分离在兔血清中生长以模拟血流感染期间遇到的环境信号的表皮细胞0-47。在2D分离后，将蛋白质转移到硝酸纤维素中，并用用活S. epidermidis 0-47或从兔血清中生长的细菌表面洗脱的生物素标记的血清蛋白。29种免疫反应性或血清结合蛋白。通过质谱鉴定表皮。其中27个基因在大肠杆菌中表达，纯化的重组蛋白用于免疫小鼠。在初步筛选中，27种重组蛋白中的12种诱导了一种反应，减少了从感染小鼠的脾脏或血流中回收的细菌数量。在随后的疫苗接种研究中，12种蛋白质中的5种导致细菌数量在统计学上显著减少。从仅29种蛋白质的初始筛选中鉴定出5种候选疫苗抗原证明了这种方法的实用性。
Staphylococcus epidermidis is a commensal of human skin and a leading cause of nosocomial bloodstream infections. Limited information is available about S. epidermidis proteins that are expressed upon transition to the bloodstream or those involved in host-pathogen interactions. A cell surface fraction from S. epidermidis 0-47 grown in rabbit serum to mimic environmental signals encountered during a bloodstream infection was separated by two-dimensional (2D) gel electrophoresis. Following 2D separation, the proteins were transferred to nitrocellulose and detected with either pooled sera generated in rabbits immunized with live S. epidermidis 0-47 or with biotin-labeled serum proteins eluted from the surface of bacteria grown in rabbit serum. Twenty-nine immunoreactive or serum binding proteins of S. epidermidis were identified by mass spectrometry. Twenty-seven of the corresponding genes were expressed in Escherichia coli, and the purified recombinant proteins were used to immunize mice. In a preliminary screen, 12 of the 27 recombinant proteins induced a response that reduced the number of bacteria recovered from the spleen or bloodstream of infected mice. In subsequent vaccination studies, 5 of the 12 proteins resulted in a statistically significant reduction in the number of bacteria. The identification of five candidate vaccine antigens from the initial screen of only 29 proteins demonstrates the utility of this approach.