Type IV Pilins as Vaccines against Bioterrorism Threats

描述(申请人提供)：IV型菌毛是许多革兰氏阴性细菌病原体的基本毒力因素，IV型菌毛蛋白已制成有效的兽用疫苗。马来伯克霍尔德氏菌和拟马来伯克霍尔德氏菌分别是扁桃体和类鼻疽病的病原体，构成了重大的生物威胁/生物恐怖主义威胁。对这些微生物未完成的基因组进行搜索后发现，它们含有相同的IV型菌毛蛋白基因。我们计划进行初步研究，为这些生物的IV型菌毛蛋白用作疫苗的未来研究提供初步数据。我们的第一个特定目标是提纯可溶性菌毛蛋白，制备抗血清，并检测实验性腺体感染后不同时间处死动物的血清，以检测对菌毛蛋白的抗体反应。我们的第二个目标是构建一株缺失菌毛蛋白基因的马来芽胞杆菌，并在体外测试其表达菌毛蛋白、菌毛及其相关表型的能力。这些研究中产生的数据和试剂将用于未来的研究，以评估IV型菌毛在实验性腺体感染中的作用，以及被动和主动菌毛免疫对腺体和类鼻疽病的保护效果。

DESCRIPTION (provided by applicant): Type IV pili are essential virulence factors for many Gram-negative bacterial pathogens and type IV pilin proteins have made effective vaccines for veterinary use. Burkholderia mallei and B. psuedomallei, the causative agents of glanders and melioidosis, respectively, represent significant biowarfare/bioterrorism threats. A search of the unfinished genomes of these microorganisms reveals that they contain identical genes for type IV pilin proteins. We plan to conduct pilot studies that will provide preliminary data for future studies of the use of the type IV pilin proteins of these organisms as vaccines. Our first specific aim is to purify soluble pilin protein, raise antisera, and test sera from animals sacrificed at various time periods after experimental glanders infection for antibody responses to pilin. Our second aim is to construct a strain of B. mallei that has a deletion in the pilin gene and test its ability to express pilin, pili and associated phenotypes in vitro. The data and reagents generated in these studies will be used for future studies to assess the role of the type IV pilus in experimental glanders infection and the protective efficacy of passive and active pilin immunization against glanders and melioidosis.