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

病毒学核心与其他内部NIAID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-D 68。 在过去一年，我们专注于开发针对大多数亚生态病毒或乙型冠状病毒的疫苗。我们成功地构建了MERS-CoV Spike及其亚基RBD和S1与SARS-CoV 2 Spike及其亚基蛋白串联融合的杂交免疫原。杂交免疫原在一个蛋白质中含有两种病毒的两个表位。此外，我们还尝试将靶免疫原蛋白负载在各种VLP上以形成潜在的更具免疫原性的候选疫苗。在动物中测试纯化的杂合蛋白、加载的VLP和编码这些免疫原的质粒DNA。 用于评估候选药物效力的小动物研究正在积极进行中。另一轮的设计和修改正在进行中，不久将在动物模型中对修改后的疫苗设计进行评估。 为了继续从免疫动物中分离高效价和广泛的抗冠状病毒单克隆抗体（mAb），我们正在使用特异性刺突亚基探针（S2 P刺突，S1或RBD）从Covid-19疫苗免疫动物中筛选和克隆B细胞。已成功分离出一些特异性结合各种新冠病毒变异体和其他冠状病毒的mAb。我们将继续进行更多的筛选试验，包括测序、EM和中和COVID-19变异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。最近的一个目标是肠道病毒D 68（EV-D 68），它会导致儿童严重的呼吸道疾病。 我们证明了EV-D 68病毒样颗粒（VLP）疫苗引发针对同源和异源EV-D 68亚进化枝的保护性中和抗体。基于B1亚进化枝2014爆发株的VLP在小鼠中引起与灭活病毒颗粒疫苗相当的B1 EV-D 68中和活性。我们的研究结果表明，疫苗株和佐剂的选择是提高对EV-D 68的保护性免疫广度的关键因素。

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.