Pre-clinical Vaccine Development for Emerging and Re-emerging Infectious Diseases

针对新发和再发传染病的临床前疫苗开发

• 批准号: 10928626
• 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/10928626
• 关键词: 2019-nCoV; Adjuvant; Animal Model; Animals; Antibodies; Antigens; B-Lymphocytes; Binding; Biological Assay; COVID-19; COVID-19 vaccine; Child; Cloning; Collaborations; Coronavirus; Coronavirus Infections; Coronavirus spike protein; Disease Outbreaks; Elements; Emerging Communicable Diseases; Enterovirus 68; Epitopes; Evaluation; Human; Hybrids; Immunity; Immunize; Middle East Respiratory Syndrome Coronavirus; Modification; Monoclonal Antibodies; Mus; National Institute of Allergy and Infectious Disease; Protein Subunits; Proteins; SARS-CoV-2 variant; Subunit Vaccines; Testing; Vaccine Design; Vaccines; Variant; Viral; Viral load measurement; Virus; Virus-like particle; betacoronavirus; coronavirus vaccine; design; hybrid protein; immunogenic; improved; neutralizing antibody; particle; pathogen; plasmid DNA; pre-clinical; respiratory; screening; vaccine candidate; vaccine development; vaccine platform; virology

The Virology Core has collaborated with other intramural NIAID labs help to develop a pan-coronaviruses spike protein subunit vaccine and a re-emerging infectious diseases, EV-D68. During the past year, we focused on developing the vaccine vaccines against most subecoviruses or beta-coronaviruses. We successfully constructed hybrid immunogens of the MERS-CoV Spike or the subunits of the spike, RBD or S1, fused with SARS-CoV2 spike or its subunit proteins tandemly. The hybrid immunogens contained both epitopes of the two viruses in one protein. In addition, we also tried to load the target immunogen proteins on various VLPs to form potentially more immunogenic vaccine candidates. The purified hybrid proteins, loaded VLPs and the plasmid DNA encoding those immunogens were tested in animals. Small animal studies for evaluating the potency of the candidates are actively on-going. Another round of designs and modification are on-going and evaluation of the modified vaccine designs in animal models will perform soon. For continuing isolation of the high potency and breadth of anti-Coronavirus monoclonal antibody (mAb) from immunized animals, we are using specific spike subunit probes (S2P spike, S1 or RBD) for screening and cloning B cells from Covid-19 vaccine immunized animals. Some mAbs specific binding to various Covid-19 emerging variants and other coronaviruses have been successfully isolated. We continue doing more screening assays, including sequencing, EM and pseudoviruses neutralizing of covid-19 variants and different coronaviruses to characterize all those mAbs. In addition, we involved in vaccine development of re-emerging infectious diseases. One recent target is the enterovirus D68 (EV-D68) which causes severe respiratory illness in children. we demonstrate that EV-D68 virus-like particle (VLP) vaccines elicit a protective neutralizing antibody against homologous and heterologous EV-D68 subclades. VLP based on a B1 subclade 2014 outbreak strain elicited comparable B1 EV-D68 neutralizing activity as an inactivated viral particle vaccine in mice. Our results suggest that both vaccine strain and adjuvant selection are critical elements for improving the breadth of protective immunity against EV-D68.

病毒学核心与 NIAID 的其他校内实验室合作，帮助开发泛冠状病毒刺突蛋白亚单位疫苗和重新出现的传染病 EV-D68。 在过去的一年里，我们专注于开发针对大多数亚生态病毒或β冠状病毒的疫苗。我们成功构建了 MERS-CoV 刺突或其亚基 RBD 或 S1 的混合免疫原，与 SARS-CoV2 刺突或其亚基蛋白串联融合。混合免疫原在一种蛋白质中含有两种病毒的两个表位。此外，我们还尝试将目标免疫原蛋白加载到各种VLP上，以形成潜在更具免疫原性的候选疫苗。纯化的杂合蛋白、负载的 VLP 和编码这些免疫原的质粒 DNA 在动物身上进行了测试。 用于评估候选药物效力的小动物研究正在积极进行中。新一轮的设计和修改正在进行中，并且很快将在动物模型中对修改后的疫苗设计进行评估。 为了持续从免疫动物中分离出高效力和广度的抗冠状病毒单克隆抗体（mAb），我们使用特定的刺突亚基探针（S2P刺突、S1或RBD）来筛选和克隆来自Covid-19疫苗免疫动物的B细胞。一些与各种 Covid-19 新变种和其他冠状病毒特异性结合的单克隆抗体已被成功分离。我们继续进行更多的筛选分析，包括测序、EM 和中和 covid-19 变体和不同冠状病毒的假病毒，以表征所有这些 mAb。 此外，我们还参与了新发传染病的疫苗开发。最近的目标之一是肠道病毒 D68 (EV-D68)，它会导致儿童严重呼吸道疾病。 我们证明 EV-D68 病毒样颗粒 (VLP) 疫苗可引发针对同源和异源 EV-D68 亚支的保护性中和抗体。基于 2014 年爆发的 B1 亚进化株的 VLP 在小鼠体内引发了与灭活病毒颗粒疫苗相当的 B1 EV-D68 中和活性。我们的结果表明，疫苗株和佐剂选择都是提高针对 EV-D68 的保护性免疫广度的关键因素。