Kudva Indira T, Griffin Robert W, Garren Jeonifer M, Calderwood Stephen B, John Manohar
Division of Infectious Diseases, Massachusetts General Hospital, Boston, MA 02114, USA.
Infect Immun. 2005 Sep;73(9):5685-96. doi: 10.1128/IAI.73.9.5685-5696.2005.
We identified spore targets of Anthrax Vaccine Adsorbed (AVA)-induced immunity in humans by screening recombinant clones of a previously generated, limited genomic Bacillus anthracis Sterne (pXO1(+), pXO2(-)) expression library of putative spore surface (spore-associated [SA]) proteins with pooled sera from human adults immunized with AVA (immune sera), the anthrax vaccine currently approved for use by humans in the United States. We identified 69 clones that reacted specifically with pooled immune sera but not with pooled sera obtained from the same individuals prior to immunization. Positive clones expressed proteins previously identified as localized on the anthrax spore surface, proteins highly expressed during spore germination, orthologs of proteins of diverse pathogens under investigation as drug targets, and orthologs of proteins contributing to the virulence of both gram-positive and gram-negative pathogens. Among the reactive clones identified by this immunological screen was one expressing a 15.2-kDa hypothetical protein encoded by a gene with no significant homology to sequences contained in databases. Further studies are required to define the subset of SA proteins identified in this study that contribute to the virulence of this pathogen. We hypothesize that optimal delivery of a subset of SA proteins identified by such studies to the immune system in combination with protective antigen (PA), the principal immunogen in AVA, might facilitate the development of defined, nonreactogenic, more-efficacious PA-based anthrax vaccines. Future studies might also facilitate the identification of SA proteins with potential to serve as targets for drug design, spore inactivation, or spore detection strategies.
我们通过用美国目前批准用于人类的炭疽疫苗吸附剂（AVA）免疫的成人混合血清（免疫血清）筛选先前构建的、有限基因组的炭疽芽孢杆菌斯特恩株（pXO1(+)，pXO2(-)）假定芽孢表面（芽孢相关[SA]）蛋白表达文库的重组克隆，确定了AVA诱导的人体免疫的芽孢靶点。我们鉴定出69个与混合免疫血清特异性反应但不与免疫前从同一受试者获得的混合血清反应的克隆。阳性克隆表达的蛋白先前被确定定位于炭疽芽孢表面，在芽孢萌发期间高表达的蛋白，作为药物靶点正在研究的多种病原体蛋白的直系同源物，以及对革兰氏阳性和革兰氏阴性病原体毒力有贡献的蛋白的直系同源物。在通过这种免疫筛选鉴定出的反应性克隆中，有一个表达由一个与数据库中序列无显著同源性的基因编码的15.2 kDa假定蛋白。需要进一步研究来确定本研究中鉴定出的有助于该病原体毒力的SA蛋白亚群。我们假设，将通过此类研究鉴定出的一部分SA蛋白与AVA中的主要免疫原保护性抗原（PA）联合最佳递送至免疫系统，可能有助于开发确定的、无反应原性的、更有效的基于PA的炭疽疫苗。未来的研究也可能有助于鉴定有潜力作为药物设计、芽孢灭活或芽孢检测策略靶点的SA蛋白。
