"Subunit Vaccines for Brucella Pathogens"

描述（由申请人提供）：布鲁氏菌属属B类制剂，是一种具有高度传染性的革兰氏阴性菌，已被确定为潜在的生物武器。布鲁氏菌病通过摄入未经巴氏消毒的乳制品自然传播，表现为流感样系统，尽管进行了抗生素治疗，但可引起反复发作的后遗症，明显表现为波状热和关节炎。布鲁氏菌在宿主体内的存活与其抵抗细胞内识别的能力有关，因此，它们可以隔离在各种组织中。迄今为止，人类还没有有效的疫苗，但保护作用是细胞介导的免疫依赖性的，特别是涉及TNF-α和IFN-γ。能够重现这种反应的疫苗应被证明对解决布鲁氏菌感染是有效的。两种推定的多胺结合蛋白，PotD和PotF，当与ISCOMs和CpG配制时，在被毒力强的melitensis 16M菌株肠外攻击的小鼠中，具有相当于Rev-1活布鲁氏菌疫苗的免疫保护作用。这种保护水平（组织定植减少bbbb4 log）尚未通过亚单位疫苗方法得到证实。鉴于这种疫苗制剂的效力，我们有独特的条件来测试这些疫苗对肠外和肺内梅氏白杆菌、流产白杆菌和猪白杆菌的有效性。因此，我们假设一种适当配方的疫苗，由pod和pof组成，结合一种适合人类使用的佐剂，将赋予预防肠外和肺布鲁氏菌攻击的保护。确定多胺结合蛋白作为疫苗靶点，最终可进一步适用于其他A类和B类病原体。为了进一步努力，特异性目标1的研究将优化布鲁氏菌疫苗（PotD和PotF）的剂量和途径，以赋予对肠外和肺B. melitensis攻击的保护，并开发保护性免疫的相关因素。特异性目标2的研究将优化佐剂与pod和pof联合使用的剂量，以获得最佳的保护，以抵御肠外和雾化的猪芽孢杆菌、流产芽孢杆菌和猪芽孢杆菌的攻击。特异性目标3的研究将在山羊布鲁氏菌病动物模型中测试PotD和PotF疫苗的功效，以确定它们是否有能力保护山羊免受黏膜挑战，防止白杆菌的定植和流产。特异性目标4的研究将评估GLP制备的pod和pof亚单位疫苗的各种安全性参数。因此，这些研究将表明，亚单位疫苗在适当提供时，可以预防肠外和肺部布鲁氏菌的挑战。

DESCRIPTION (provided by applicant): Brucella species, members of Category B agents, are highly infectious Gram-negative bacteria that have been identified as potential biological weapons. Brucellosis is naturally transmitted via ingestion of unpasteurized dairy products, manifesting with flu-like systems and, despite antibiotic treatment, can cause a recurring sequelae, evident as undulant fever and arthritis. Brucellae survival within the host is linked to its ability to resist intracellular recognition, thus, allowing them to sequester in various tissues. To date, there are no effective vaccines for humans, but protection is cell-mediated immunity-dependent, particularly, involving TNF-α and IFN-γ. Vaccines that can recapitulate such responses should prove effective in resolving Brucella infections. Two putative polyamine-binding proteins, PotD and PotF, when formulated with ISCOMs plus CpG, conferred immune protection equivalent to the live Rev-1 Brucella melitensis vaccine in mice parenterally challenged with virulent B. melitensis 16M strain. This level of protection (>4 log reduction in tissue colonization) has yet to be shown using a subunit vaccine approach. Given the potency of this vaccine formulation, we are uniquely poised to test the efficacy of these vaccines against parenteral and pulmonary B. melitensis, B. abortus, and B. suis challenges. Thus, we hypothesize that an appropriately formulated vaccine composed of PotD and PotF, combined with a suitable adjuvant for human use, will confer protection against parenteral and pulmonary Brucella challenge. Identifying polyamine-binding proteins as vaccine targets could eventually be further adapted for other Category A and B pathogens. To further this effort, studies in Specific Aim 1 will optimize the dosage and route for Brucella vaccines, PotD and PotF, to confer protection against parenteral and pulmonary B. melitensis challenges and develop correlates for protective immunity. Studies in Specific Aim 2 will optimize the dosage for adjuvant when combined with PotD and PotF to confer optimal protection against parenteral and aerosolized B. melitensis, B. abortus, and B. suis challenges. Studies in Specific Aim 3 will test the efficacy of PotD and PotF vaccines in a caprine brucellosis animal model for their ability to confer protection against mucosally challenged goats for B. melitensis colonization and abortion. Studies in Specific Aim 4 will evaluate various safety parameters for GLP- prepared PotD and PotF subunit vaccines. Thus, these studies will show that subunit vaccines, when appropriately delivered, can protect against parenteral and pulmonary Brucella challenges.