Delvecchio Vito G, Connolly Joseph P, Alefantis Timothy G, Walz Alexander, Quan Marian A, Patra Guy, Ashton John M, Whittington Jessica T, Chafin Ryan D, Liang Xudong, Grewal Paul, Khan Akbar S, Mujer Cesar V
Vital Probes, Inc., 1300 Old Plank Road, Mayfield, PA 18433, USA.
Appl Environ Microbiol. 2006 Sep;72(9):6355-63. doi: 10.1128/AEM.00455-06.
Differentially expressed and immunogenic spore proteins of the Bacillus cereus group of bacteria, which includes Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis, were identified. Comparative proteomic profiling of their spore proteins distinguished the three species from each other as well as the virulent from the avirulent strains. A total of 458 proteins encoded by 232 open reading frames were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry analysis for all the species. A number of highly expressed proteins, including elongation factor Tu (EF-Tu), elongation factor G, 60-kDa chaperonin, enolase, pyruvate dehydrogenase complex, and others exist as charge variants on two-dimensional gels. These charge variants have similar masses but different isoelectric points. The majority of identified proteins have cellular roles associated with energy production, carbohydrate transport and metabolism, amino acid transport and metabolism, posttranslational modifications, and translation. Novel vaccine candidate proteins were identified using B. anthracis polyclonal antisera from humans postinfected with cutaneous anthrax. Fifteen immunoreactive proteins were identified in B. anthracis spores, whereas 7, 14, and 7 immunoreactive proteins were identified for B. cereus and in the virulent and avirulent strains of B. thuringiensis spores, respectively. Some of the immunodominant antigens include charge variants of EF-Tu, glyceraldehyde-3-phosphate dehydrogenase, dihydrolipoamide acetyltransferase, Delta-1-pyrroline-5-carboxylate dehydrogenase, and a dihydrolipoamide dehydrogenase. Alanine racemase and neutral protease were uniquely immunogenic to B. anthracis. Comparative analysis of the spore immunome will be of significance for further nucleic acid- and immuno-based detection systems as well as next-generation vaccine development.
已鉴定出蜡样芽孢杆菌属细菌（包括炭疽芽孢杆菌、蜡样芽孢杆菌和苏云金芽孢杆菌）中差异表达且具有免疫原性的芽孢蛋白。对它们芽孢蛋白的比较蛋白质组分析区分了这三个物种，以及有毒菌株和无毒菌株。通过基质辅助激光解吸电离飞行时间质谱分析，鉴定出了所有物种中由232个开放阅读框编码的总共458种蛋白质。许多高表达蛋白，包括延伸因子Tu（EF-Tu）、延伸因子G、60 kDa伴侣蛋白、烯醇化酶、丙酮酸脱氢酶复合体等，在二维凝胶上以电荷变体形式存在。这些电荷变体具有相似的质量但不同的等电点。大多数鉴定出的蛋白质具有与能量产生、碳水化合物运输和代谢、氨基酸运输和代谢、翻译后修饰以及翻译相关的细胞功能。使用来自皮肤炭疽感染后人类的炭疽芽孢杆菌多克隆抗血清鉴定出了新型疫苗候选蛋白。在炭疽芽孢杆菌芽孢中鉴定出了15种免疫反应性蛋白，而在蜡样芽孢杆菌以及苏云金芽孢杆菌有毒和无毒菌株的芽孢中分别鉴定出了7种和14种以及7种免疫反应性蛋白。一些免疫显性抗原包括EF-Tu的电荷变体、甘油醛-3-磷酸脱氢酶、二氢硫辛酰胺乙酰转移酶、Δ-1-吡咯啉-5-羧酸脱氢酶和一种二氢硫辛酰胺脱氢酶。丙氨酸消旋酶和中性蛋白酶对炭疽芽孢杆菌具有独特的免疫原性。芽孢免疫组的比较分析对于进一步基于核酸和免疫的检测系统以及下一代疫苗开发具有重要意义。
