Production of an NP-ISS conjugate vaccine to combat pandemic influenza.

描述（由申请方提供）：本提案的目的是继续开发一种新型疫苗方法的产品，该方法将核蛋白（NP）作为流感病毒的保守内部组分与免疫刺激寡核苷酸连接，以诱导针对不同和潜在大流行性流感毒株的保护性免疫。具体而言，该计划是完成工艺开发并在E. coli系统。然后将进行疫苗用NP-ISS结合物的工艺开发、放大和GMP生产，以及IND毒理学和稳定性研究。此外，将在动物模型中对NP-ISS疫苗单独接种和NP-ISS与标准三价灭活疫苗联合接种提供的免疫原性和保护性免疫进行表征。该提案的具体目标是： 1.生产GMP级NP-ISS结合物 * 重组核蛋白在大肠杆菌中的GMP生产。大肠杆菌生产系统 * NP-ISS结合物制剂的工艺开发和放大 * NP-ISS制剂的GMP生产 2.对NP-ISS疫苗<$TIV进行IND启用毒理学 3.对NP-ISS疫苗进行进一步表征。 * 评估E提供的保护。在针对漂移（H1）、转变（H3）和禽（H5）流感毒株的小鼠攻击模型中， * 在雪貂流感攻毒模型中，使用漂移（H1）、转换（H3）和禽（H5）流感毒株评估免疫后的保护作用。 * 在灵长类动物中优化NP-ISS和TIV的组合，以允许HA剂量节省，同时诱导NP特异性细胞介导的免疫。 * 评价NP-ISS疫苗联合三价灭活疫苗在老年小鼠中的免疫原性。

DESCRIPTION (provided by applicant): The goal of this proposal is to continue the product development of a novel vaccine approach that incorporates nucleoprotein (NP) as a conserved internal component of influenza virus linked to an immunostimulatory oligonucleotide to induce protective immunity against divergent and potentially pandemic influenza strains. Specifically, the plan is to complete process development and scale up for GMP production of recombinant NP in an E. coli system. Process development, scale-up and GMP production of the NP-ISS conjugate for the vaccine will then be performed, as well as IND enabling toxicology and stability studies. In addition, characterization of the immunogenicity and protective immunity provided by the NP-ISS vaccine alone and NP-ISS combined with standard trivalent inactivated vaccine will be performed in animal models. The Specific Aims of the proposal are: 1. Produce GMP-grade NP-ISS conjugate * GMP manufacture of recombinant nucleoprotein in the E. coli production system * Process development and scale-up of NP-ISS conjugate formulation * GMP manufacture of NP-ISS formulation 2. Perform IND enabling toxicology on NP-ISS vaccine ¿ TIV 3. Perform further characterization of the NP-ISS vaccine. * Assess protection provided by E. coli derived NP-ISS in the mouse challenge model against drift (H1), shift (H3) and avian (H5) influenza strains. * Assess protection following immunization in the ferret influenza challenge model with drift (H1),shift (H3) and avian (H5) influenza strains. * Optimize the combination of NP-ISS and TIV in primates to allow HA dose sparing while inducing NP-specific cell mediated immunity. * Evaluate immunogenicity of the NP-ISS vaccine combined with trivalent inactivated vaccine in elderly mice.