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

描述(由申请人提供)：这项建议的目标是继续开发一种新的疫苗方法，将核蛋白(NP)作为流感病毒的保守内部成分与免疫刺激寡核苷酸相连，以诱导对不同的和潜在的大流行流感毒株的保护性免疫。具体地说，该计划是完成工艺开发和扩大规模，在大肠杆菌系统中生产重组NP的GMP。随后将进行疫苗用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单独疫苗和NP-ISS与标准三价灭活疫苗联合提供的免疫原性和保护性免疫进行表征。这项建议的具体目标是： 1.生产GMP级NP-ISS结合物 *在大肠杆菌生产系统中生产重组核蛋白的GMP *NP-ISS共轭配方的工艺开发和放大 *生产NP-ISS配方的GMP 2.对NP-ISS疫苗进行IND启用毒理学？TIV 3.对NP-ISS疫苗进行进一步表征。 *评估大肠杆菌衍生的NP-ISS在小鼠挑战模型中对漂移(H1)、移位(H3)和禽流感(H5)病毒株的保护作用。 *评估雪貂流感挑战模型中使用漂移(H1)、移位(H3)和禽流感(H5)毒株接种后的保护情况。 *优化灵长类动物中NP-ISS和TIV的组合，以便在诱导NP特异性细胞免疫的同时节省HA剂量。 *评估NP-ISS疫苗与三价灭活疫苗联合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.