Preclinical Development of Flagellin as a Mucosal Adjuvant in Biodefense Vaccines

描述（由申请人提供）：革兰氏阴性鞭毛蛋白是先天性和适应性免疫应答的非常有效的激活剂。我们和其他人的工作已经确定鞭毛蛋白是小鼠的有效佐剂。我们还发现鞭毛蛋白在非人灵长类动物中是一种有效的佐剂。鞭毛蛋白具有使其成为优良佐剂的特征，包括：效力;粘膜活性;在鞭毛蛋白免疫宿主中的有效性;易于在细菌中产生;和5）在蛋白质的高变区中插入抗原序列而不损失鞭毛蛋白生物活性的能力。我们已经清楚地证实，鞭毛蛋白是一种非常有效的粘膜佐剂，在发展一个强大的，特异性的抗体反应，对Y。在小鼠和非人灵长类动物中的鼠疫抗原，也促进对致命的呼吸道攻击的保护与Y。鼠疫由于鞭毛蛋白具有作为用于人类的粘膜佐剂的巨大潜力，我们提出了以下目标，这将促进鞭毛蛋白用于人类用途：具体目标1。确定鞭毛蛋白/Y.鼠疫F1和V融合蛋白作为保护性粘膜疫苗。我们已经产生了一个重组蛋白编码鞭毛蛋白和F1和V抗原的Y。鼠疫杆菌的融合蛋白，并且已经确定该融合蛋白保留鞭毛蛋白生物活性。我们建议确定融合蛋白是否促进小鼠和食蟹猴的抗F1和V IgG应答以及小鼠对Y.鼠疫CO 92。具体目标2。设计并实施符合药品生产质量管理规范（GMP）的鞭毛蛋白/Y生产工艺。鼠疫疫苗，可以很容易地扩大规模，用于临床前，临床和最终的商业用途。具体目标3。使用鞭毛蛋白/Y进行GLP毒理学研究。鼠疫疫苗工艺优化后，将制备工程批次，以在GLP条件下在小鼠模型中研究其安全性和免疫原性。该批次还将用于表征研究，并启动正式稳定性研究，为临床生产做准备。具体目标4。为了产生鞭毛蛋白/Y.在良好生产规范（GMP）条件下生产鼠疫疫苗。具体目标2中制定的方案将用于在GMP条件下生产克量疫苗。这些目标的实现将为粘膜鞭毛蛋白/Y的临床评价奠定坚实的基础。人类的鼠疫疫苗

DESCRIPTION (provided by applicant): Gram-negative flagellin is an extraordinarily potent activator of innate and adaptive immune responses. Our work as well as that of others has established that flagellin is a potent adjuvant in mice. We have also found that flagellin is a potent adjuvant in non-human primates. Flagellin possesses features that make it an excellent adjuvant including: potency; mucosally active; effectiveness in flagellin immune hosts; ease of production in bacteria; and 5) the ability to insert antigen sequences in the hypervariable region of the protein without loss of flagellin bioactivity. We have clearly established that flagellin is a highly effective mucosal adjuvant in the development of a strong, specific antibody response against Y. pestis antigens in mice and non-human primates and also promotes protection against a lethal respiratory challenge with Y. pestis. Since flagellin possesses tremendous potential as a mucosal adjuvant for use in humans, we propose the following aims that will advance the development of flagellin for human use: Specific Aim 1. To determine the efficacy of a flagellin/Y. pestis F1 and V fusion protein as a protective mucosal vaccine. We have generated a recombinant protein encoding flagellin and both the F1 and V antigens of Y. pestis and have established that this fusion protein retains flagellin biological activity. We propose to determine if the fusion protein promotes a robust anti-F1 and V IgG response in mice and cynomolgus monkeys and protection in mice against respiratory challenge with Y. pestis CO92. Specific Aim 2. To design and implement a Good Manufacturing Practice (GMP) compatible production process for a flagellin/Y. pestis vaccine that can be readily scaled for pre-clinical, clinical and ultimately commercial use. Specific Aim 3. To conduct GLP toxicology studies with the flagellin/Y. pestis vaccine. Once the process has been optimized, an engineering lot will be prepared to study its safety and immunogenicity under GLP conditions in a mouse model. This lot will also be used for characterization studies and to initiate formal stability studies in preparation for clinical manufacture. Specific Aim 4. To produce a flagellin/Y. pestis vaccine under Good Manufacturing Practices (GMP) conditions. The protocol developed in Specific Aim 2 will be used to produce gram quantities of the vaccine under GMP conditions. The completion of these aims will provide a very strong foundation for the clinical evaluation of the mucosal flagellin/Y. pestis vaccine in humans.