Eyles Jim E, Unal Berkay, Hartley Margaret Gill, Newstead Sarah L, Flick-Smith Helen, Prior Joann L, Oyston Petra C F, Randall Arlo, Mu Yunxiang, Hirst Siddiqua, Molina Douglas M, Davies D Huw, Milne Tim, Griffin Kate F, Baldi Pierre, Titball Rick W, Felgner Philip L
Defence Science and Technology Laboratory, Porton Down, Salisbury, UK.
Proteomics. 2007 Jun;7(13):2172-83. doi: 10.1002/pmic.200600985.
Stimulation of protective immune responses against intracellular pathogens is difficult to achieve using non-replicating vaccines. BALB/c mice immunized by intramuscular injection with killed Francisella tularensis (live vaccine strain) adjuvanted with preformed immune stimulating complexes admixed with CpG, were protected when systemically challenged with a highly virulent strain of F. tularensis (Schu S4). Serum from immunized mice was used to probe a whole proteome microarray in order to identify immunodominant antigens. Eleven out of the top 12 immunodominant antigens have been previously described as immunoreactive in F. tularensis. However, 31 previously unreported immunoreactive antigens were revealed using this approach. Twenty four (50%) of the ORFs on the immunodominant hit list belonged to the category of surface or membrane associated proteins compared to only 22% of the entire proteome. There were eight hypothetical protein hits and eight hits from proteins associated with different aspects of metabolism. The chip also allowed us to readily determine the IgG subclass bias, towards individual or multiple antigens, in protected and unprotected animals. These data give insight into the protective immune response and have potentially important implications for the rational design of non-living vaccines for tularemia and other intracellular pathogens.
使用非复制型疫苗很难激发针对细胞内病原体的保护性免疫反应。用预先形成的免疫刺激复合物与CpG混合佐剂的灭活土拉弗朗西斯菌（活疫苗株）通过肌肉注射免疫BALB/c小鼠，当用高毒力的土拉弗朗西斯菌（Schu S4）株进行全身攻击时，这些小鼠受到了保护。用免疫小鼠的血清探测全蛋白质组微阵列以鉴定免疫显性抗原。前12种免疫显性抗原中有11种先前已被描述为在土拉弗朗西斯菌中具有免疫反应性。然而，使用这种方法发现了31种先前未报道的免疫反应性抗原。免疫显性命中列表上24个（50%）的开放阅读框属于表面或膜相关蛋白类别，而整个蛋白质组中这一比例仅为22%。有8个假定蛋白命中和8个来自与代谢不同方面相关的蛋白命中。该芯片还使我们能够轻松确定受保护和未受保护动物中针对单个或多个抗原的IgG亚类偏向。这些数据深入了解了保护性免疫反应，对兔热病和其他细胞内病原体的非活性疫苗的合理设计具有潜在的重要意义。
