Novel vaccine against Norovirus

描述（由申请人提供）：辛辛那提儿童医院医学中心（CCHMC）、弗吉尼亚理工学院和州立大学（弗吉尼亚理工大学）以及LigoCyte制药公司将根据RFA-AI-09-027，题为“生物防御食品和水传播疾病的合作伙伴关系（R01）”的申请进行合作。我们建议以新发现的诺如病毒的亚病毒粒子P粒子为基础，研制一种针对诺如病毒（NVs）的疫苗。诺如病毒是NIAID B类食物和水传播的优先病原体之一。该P颗粒由病毒衣壳蛋白的24个突起(P)结构域自发形成，呈八面体对称，具有真实的宿主受体结合特性，具有高度的免疫原性，易于在大肠杆菌和酵母中产生，产量极高，纯化程序简单，在广泛的物理化学条件下高度稳定，是一种优良的粘膜免疫亚基疫苗。将进行四个项目，代表不同的早期到中期产品开发阶段，以评估P颗粒疫苗。在第一阶段，我们将进行体外和小鼠免疫研究，并比较不同途径的P颗粒的不同疫苗接种方案，以获得最大的安全性和有效性。我们还将使用新发现的猴杯状病毒（Tulane病毒）的细胞培养模型进行类似的研究，以证明P颗粒疫苗的有用性。在第二阶段，我们将描述代表nv不同基因型的nv的抗原变异特征，以便根据确定的抗原型（血清型），采用鸡尾酒或共识疫苗方法开发具有广泛反应性的疫苗。我们还将对目前占主导地位的GII-4病毒进行遗传和抗原分析，以解决nv的划时代进化问题，为未来针对nv的疫苗策略提供帮助。在第三阶段，我们将利用我们实验室最近开发的猪对人类nv的非生物攻击模型，进一步表征候选疫苗在预防各种nv方面的安全性、免疫原性和广泛性。在最后阶段，我们将开发一个发酵生物反应器，以扩大P颗粒的生产规模，以便将来通过人体志愿者挑战研究和临床试验对疫苗进行评估。nv具有遗传多样性，没有合适的细胞培养物。我们的鸡尾酒/共识疫苗方法以及通过替代“中和”试验和使用新发现的杜兰病毒和Gn猪模型对疫苗进行评估的程序具有高度创新性。我们有信心，通过本申请的研究结论，将选出一组主要候选（疫苗株），为未来的临床试验做好准备。

DESCRIPTION (provided by applicant): The Cincinnati Children's Hospital Medical Center (CCHMC), the Virginia Polytechnic Institute and State University (the Virginia Tech), and the LigoCyte Pharmaceuticals Inc. will team together for this application in response to RFA-AI-09-027, entitled "Partnerships for Biodefense Food- and Water-borne Diseases (R01). We propose to develop a vaccine against noroviruses (NVs), one of the NIAID Category B food- and water-borne priority pathogens, based on our newly discovered subviral particle, the P particle, of NVs. This P particle is spontaneously formed by 24 copies of the protrusion (P) domain of the viral capsid protein, in octahedral symmetry, with an authentic host receptor binding property, highly immunogenic, easily to produce in E. coli and yeast with extremely high yields and simple procedures for purification, and highly stable under a wide range of physic-chemical conditions, making it an excellent subunit vaccine for mucosal immunization. Four projects representing different early-to-middle product development stages will be performed to evaluate the P particle vaccine. In stage 1 we will perform in vitro and mouse immunization studies and compare different vaccination regimens of P particles by different routes with/without an adjuvant for a maximal safety and efficacy. We also will perform similar studies for proof-of-concept of the usefulness of P particle vaccine using the cell culture model of a newly discovered monkey calicivirus, the Tulane virus. In stage 2 we will characterize the antigenic variations of NVs representing different genotypes of NVs for developing a broadly reactive vaccine based on defined antigenic types (serotypes) with a cocktail or consensus vaccine approach. We also will perform genetic and antigenic analysis of the currently dominant GII-4 viruses to address a question about epochal evolution of NVs for future vaccine strategy against NVs. In stage 3 we will further characterize the safety, immunogenicity and broadness of the candidate vaccine in protection against various NVs using the gnotobiotic (Gn) pig challenge model of human NVs developed in our laboratories recently. In the last stage we will develop a fermentation bioreactor for scale up production of P particles for future evaluation of the vaccine by human volunteer challenge studies and clinical trials. NVs are genetically diverse and there is no suitable cell culture for NVs. Our cocktail/consensus vaccine approach and the procedures for evaluation of the vaccine by the surrogate "neutralization" assay and using the newly discovered Tulane virus and the Gn pig model are highly innovative. We are confident that a panel of lead candidates (vaccine strains) will be selected ready for future clinical trials by the conclusion of the studies in this application.