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冠状病毒暴发引发全球健康担忧时，我们启动了一项开发候选疫苗的计划。我们从S的S蛋白序列出发，建立了一种由S全长DNA基元和S1亚基蛋白Boost组成的免疫策略，可诱导针对8种不同MERS冠状病毒株的高滴度中和抗体。免疫血清含有针对受体结合域(RBD)、S1的非RBD部分和S2亚单位的有效中和抗体。从免疫的小鼠中，我们产生了一组杂交瘤和产生的单抗，从中选择了一种具有高中和活性的品种进行进一步的鉴定。与RBD结合的单抗D12的原子结构揭示了它们阻断与MERS冠状病毒受体DPP4结合的两种不同机制。此外，还测量了非人灵长类动物的免疫原性。因此，由S序列设计的疫苗免疫原诱导了不同的中和抗体库，展示了一种可能适用于其他新兴病毒的疫苗设计的有效方法。对HKU1β冠状病毒的刺突糖蛋白也进行了结构研究，以探索结构和发现受体，并产生了刺突三聚体结构溶液。建立人呼吸道上皮细胞培养体系，以启动HKU1的进入和致病机制研究。通过合作，我们已经解决了MERS S蛋白的结构，并在此结构的基础上设计了稳定突变，使S蛋白稳定在其预融合构象中。我们目前正在评估稳定的MERS S蛋白的免疫原性，并评估其他冠状病毒S蛋白的稳定突变。