Coronavirus vaccine development

冠状病毒疫苗开发

• 批准号: 9551285
• 负责人: Barney Graham
• 金额: $ 84.1万
• 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/9551285
• 关键词: Antigens; Binding; Camels; Cell Culture System; Collaborations; Complex; Coronaviridae; Coronavirus; Coronavirus spike protein; DNA; Disease Outbreaks; Epithelial Cells; Glycoproteins; Goals; Human; Hybridomas; Immune Sera; Immunization; Immunize; Length; Measures; Middle East Respiratory Syndrome Coronavirus; Molecular Conformation; Monoclonal Antibodies; Mus; Mutation; Pathogenesis; Protein Subunits; Proteins; Public Health; SARS coronavirus; Structure; Technology; Vaccine Design; Vaccines; Virus; base; design; global health; immunogenicity; neutralizing antibody; nonhuman primate; programs; receptor; receptor binding; vaccine candidate; vaccine development

When the MERS CoV outbreak raised global health concerns, we initiated a program to develop a candidate vaccine. Starting from Spike glycoprotein (S) sequences, we developed an immunization strategy consisting of a full-length S DNA prime and a S1 subunit protein boost that elicited high titers of neutralizing antibodies against eight different MERS-CoV strains. Immune sera contained potent neutralizing antibodies targeting the receptor binding domain (RBD), non-RBD portions of S1, and the S2 subunit. From the immunized mice, we produced a panel of hybridomas and produced monoclonal antibodies from which a variety with high neutralizing activity were selected for further characterization. The atomic structure of a monoclonal antibody, D12, in complex with the RBD revealed two distinct mechanisms by which they block binding to the MERS-CoV receptor, DPP4. In addition, immunogenicity was measured in nonhuman primates. Thus, vaccine immunogens designed from S sequences induced a diverse repertoire of neutralizing antibodies, demonstrating an efficient approach to vaccine design that may be applicable to other emerging viruses. Structural studies were also initiated with the spike glycoprotein of the HKU1 beta-coronavirus, to explore structure and for receptor discovery and led to a spike trimer structure solution. Human airway epithelial cell culture system was established to initiate entry and pathogenesis studies of HKU1. Through collaborations, we have solved the structure of the MERS S protein and based on this structure, designed stabilizing mutations which stabilize the S protein in its prefusion conformation. We are currently assessing immunogenicity of the stabilized MERS S protein, and evaluating stabilizing mutations in other coronavirus spike proteins.

当MERS COV爆发引起全球健康问题时，我们启动了一项计划来开发候选疫苗。从峰值糖蛋白序列开始，我们制定了一种免疫策略，该策略由全长S DNA素和S1亚基蛋白增强，从而引起了针对八种不同MERS-COV菌株的中和抗体的高滴度。免疫血清包含针对受体结合结构域（RBD），S1的非RBD部分和S2亚基的有效中和抗体。从免疫小鼠中，我们产生了一组杂交瘤，并产生了单克隆抗体，从中选择了具有高中和活性的种类以进一步表征。单克隆抗体D12的原子结构与RBD复合揭示了两种不同的机制，它们可以阻止与MERS-COV受体DPP4结合的结合。另外，在非人类灵长类动物中测量了免疫原性。因此，由S序列设计的疫苗免疫原诱导了中和抗体的多种曲目，证明了一种有效的疫苗设计方法，可能适用于其他新兴病毒。结构研究还使用HKU1β-核负病毒的尖峰糖蛋白开始，以探索结构和受体发现，并导致尖峰三聚体结构溶液。建立了人类气道上皮细胞培养系统以启动HKU1的进入和发病机理研究。 通过合作，我们解决了MERS蛋白的结构，并基于这种结构，设计了稳定突变，以稳定S蛋白在其预融合构象中。 我们目前正在评估稳定的MERS蛋白的免疫原性，并评估其他冠状病毒尖峰蛋白中的稳定突变。