Pre-clinical Vaccine Development for Respiratory Viruses

呼吸道病毒临床前疫苗开发

• 批准号: 10928627
• 负责人: Wing Pui Kong
• 金额: $ 248.09万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10928627
• 关键词: Antigens; Chimeric Proteins; Cryoelectron Microscopy; Discrimination; Disulfides; Elderly; Engineering; Generations; Glycoproteins; Human; Immunization; Immunocompromised Host; Individual; Infection; Macaca mulatta; Modification; Mus; Mutation; Newborn Infant; Para-Influenza Virus Type 1; Paramyxovirus; Pneumovirus; Publishing; Rhesus; Site; Structure; Surface; Vaccine Antigen; Vaccines; burden of illness; design; disulfide bond; flexibility; human pathogen; immunogenicity; nanoparticle; nonhuman primate; pathogen; pre-clinical; respiratory; respiratory virus; response; vaccine candidate; vaccine development; vaccine immunogenicity; vaccine response

Human paramyxoviruses and pneumoviruses are widespread pathogens, cause considerable disease burden. Parainfluenza virus types 1-4 (PIV1-4) are highly infectious human pathogens, of which PIV3 is most commonly responsible for severe respiratory illness in newborns, elderly, and immunocompromised individuals. Based on a cryoelectron microscopy structure of an engineered PIV3 F prefusion-stabilized trimer, we engineered in additional mutations different from our previous published modifications in each of the four PIV fusion (F) glycoproteins to further stabilize their metastable prefusion states. The vaccine candidates were able to elicit murine PIV type-specific response, with little cross-neutralization of other PIVs. We evaluated the neutralization responses of the vaccine candidates in nonhuman primates (NHPs). We applied similar concept and designs to hMPV F proteins. We designed various mutations to further stabilize the PIV F prefusion trimers and hMPV prefusion trimers. In addition, we designed expression the modified trimers on the surface of different nanoparticles as vaccine immunogens. In the past year, we investigate the impact of further stabilizing HMPV F in the pre-F state. We replaced the furin-cleavage site with a flexible linker, creating a single chain F that yielded increased amounts of pre-F stabilized trimers, enabling the generation and assessment of F trimers stabilized by multiple disulfide bonds. Immunogenicity assessments in nave mice showed the triple disulfide-stabilized pre-F trimer could elicit high titer neutralization than elicited by post-F. Immunogenicity assessments in pre-exposed rhesus macaques showed the triple disulfide-stabilized pre-F could recall high neutralizing titers after a single immunization, with little discrimination in the recall response between pre-F and post-F. Collectively, these results suggest single-chain triple disulfide-stabilized pre-F trimers to be promising HMPV-vaccine antigens.

人类副粘病毒和肺炎病毒是广泛分布的病原体，造成相当大的疾病负担。副流感病毒1-4型（PIV 1 -4）是高度传染性的人类病原体，其中PIV 3最常导致新生儿、老年人和免疫功能低下个体的严重呼吸道疾病。基于工程化PIV 3 F融合前稳定三聚体的冷冻电子显微镜结构，我们在四种PIV融合（F）糖蛋白中的每一种中设计了与我们先前发表的修饰不同的其他突变，以进一步稳定其亚稳态融合前状态。候选疫苗能够引起鼠PIV类型特异性应答，几乎没有其他PIV的交叉中和。我们评价了候选疫苗在非人灵长类动物（NHP）中的中和反应。 我们将类似的概念和设计应用于hMPV F蛋白。我们设计了各种突变以进一步稳定PIV F融合前三聚体和hMPV融合前三聚体。此外，我们设计了将修饰后的三聚体表达在不同纳米颗粒表面作为疫苗免疫原。 在过去的一年中，我们研究了进一步稳定HMPV F在预F状态的影响。 我们用柔性接头取代弗林蛋白酶切割位点，产生单链F，产生增加量的前F稳定的三聚体，使得能够产生和评估由多个二硫键稳定的F三聚体。免疫原性评估表明，三重二硫键稳定的前F三聚体可以引起高滴度的中和比后F。在预先暴露的恒河猴中进行的免疫原性评估显示，三重二硫键稳定的前F在单次免疫后可以回忆起高中和滴度，前F和后F之间的回忆反应几乎没有区别。总的来说，这些结果表明单链三重二硫键稳定的前F三聚体是有希望的HMPV疫苗抗原。