Type IV Pilins as Vaccines against Bioterrorism Threats

描述（由申请人提供）：IV型菌毛是许多革兰氏阴性细菌病原体必不可少的毒力因子，IV型菌毛蛋白已制成有效的兽医用疫苗。马氏伯克霍尔德菌和假多囊杆菌分别是腺体病和类鼻疽病的病原体，它们代表着重大的生物战/生物恐怖威胁。对这些微生物未完成的基因组的搜索显示，它们含有相同的IV型pilin蛋白基因。我们计划进行试点研究，为今后使用这些生物的IV型蛋白作为疫苗的研究提供初步数据。我们的第一个具体目标是纯化可溶性匹林蛋白，培养抗血清，并在实验腺感染后不同时期牺牲动物的血清中检测对匹林的抗体反应。我们的第二个目标是构建一株具有pilin基因缺失的mallei B.菌株，并测试其在体外表达pilin， pili和相关表型的能力。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型皮针在实验性腺体感染中的作用，以及被动和主动皮针免疫对腺体和类鼻疽的保护作用。

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.