Structure based design of dengue subunit vaccines for inducing protective but not disease enhancing antibodies

基于结构的登革热亚单位疫苗设计，用于诱导保护性而非疾病增强性抗体

• 批准号: 10612354
• 负责人: Aravinda M. DeSilva
• 金额: $ 72.32万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-21 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10612354
• 关键词: Antibodies; Antibody Response; Attenuated; B-Lymphocytes; Binding; Cell secretion; Child; Clinical Trials; Computing Methodologies; Culicidae; Dengue; Dengue Infection; Dengue Vaccine; Dengue Virus; Dengvaxia; Disease; E protein; Epitopes; Exposure to; Flavivirus; Goals; Human; Immune response; Immunity; Immunize; Infection; Length; Mammalian Cell; Membrane; Membrane Proteins; Mus; Mutate; Persons; Physiological; Placebos; Process; Production; Protein Secretion; Proteins; Recombinant Proteins; Recombinants; Research; Rest; Risk; Safety; Serotyping; Severity of illness; Specificity; Structure; Subunit Vaccines; Surface; Techniques; Testing; Vaccination; Vaccine Antigen; Vaccine Design; Vaccines; Variant; Virion; Virus; Virus Diseases; Virus Replication; Virus-like particle; biophysical properties; computerized tools; cross reactivity; design; dimer; e Antigens; env Gene Products; immunogenicity; immunological status; molecular modeling; monomer; mouse model; neutralizing antibody; novel; particle; preservation; protein folding; public trust; recombinant virus; response; severe dengue; stability testing; success; vaccination strategy; vaccine candidate; vaccine development; viral transmission; virus envelope

ABSTRACT Dengue virus (DENV) vaccine development has been challenging because of the presence of 4 serotypes (DENV1-4) and the potential for vaccine enhanced severe disease. The leading live attenuated tetravalent DENV vaccines have been plagued by poorly balanced replication of vaccine components leading to variable efficacy and vaccine primed severe dengue disease in some children. The goal of this proposal is to develop novel recombinant DENV envelope (E) protein and virus like particle (VLP) vaccines that overcome barriers faced by live attenuated tetravalent vaccines. We have discovered that the DENV E protein produced as a secreted protein is a poor vaccine because it is a monomer that does not display major quaternary epitopes targeted by human neutralizing and protective antibodies (Abs). This proposal is based on new discoveries from our group about how structure based, computational approaches can be used to produce highly stable and properly folded DENV E dimers that are efficiently secreted from mammalian cells. We will use mouse models of DENV vaccination and infection to test if artificially stabilized DENV E dimers stimulate Ab responses that are similar to serotype-specific and serotype-cross-protective Ab responses in people exposed to primary and secondary wild type DENV infections. The stabilized E dimers will be further modified to test if large-scale resurfacing of the E dimer can be used to focus the immune response on epitopes recognized by potent neutralizing Abs, while eliminating responses to off target, disease enhancing epitopes. Finally, we will design membrane anchored variants of stabilized E dimers to promote the formation of dengue virus-like particle (VLP) vaccine candidates that better resemble mature infectious virions than VLPs made with current techniques. Our studies, which explore how to design and deliver recombinant E antigens to focus the host antibody response on important quaternary structure neutralizing epitopes, will stimulate new research directions in the field of flavivirus subunit vaccines.

抽象的 由于存在 4 种血清型，登革热病毒 (DENV) 疫苗的开发一直充满挑战 （DENV1-4）和疫苗增强严重疾病的潜力。领先的减毒活四价 DENV 疫苗一直受到疫苗成分复制不平衡的困扰，导致功效参差不齐 疫苗在一些儿童中引发了严重的登革热疾病。该提案的目标是开发新颖的 重组 DENV 包膜 (E) 蛋白和病毒样颗粒 (VLP) 疫苗克服了所面临的障碍 四价减毒活疫苗。我们发现 DENV E 蛋白是作为分泌型 蛋白质是一种较差的疫苗，因为它是一种单体，不显示主要的四级表位 人类中和和保护性抗体（Abs）。该提案基于我们小组的新发现 关于如何使用基于结构的计算方法来生产高度稳定且正确折叠的产品 由哺乳动物细胞有效分泌的 DENV E 二聚体。我们将使用DENV的小鼠模型 疫苗接种和感染，以测试人工稳定的 DENV E 二聚体是否会刺激类似于 暴露于初级和次级野生动物的人群中血清型特异性和血清型交叉保护性抗体反应 DENV 型感染。稳定的E二聚体将被进一步修饰，以测试E的大规模表面重建 二聚体可用于将免疫反应集中在有效中和抗体识别的表位上，同时 消除对脱靶、疾病增强表位的反应。最后，我们将设计膜锚固 稳定的 E 二聚体变体促进登革热病毒样颗粒 (VLP) 候选疫苗的形成 与用现有技术制造的 VLP 相比，它更类似于成熟的传染性病毒体。我们的研究， 探索如何设计和递送重组 E 抗原，以将宿主抗体反应集中于重要的 四级结构中和表位，将激发黄病毒亚基领域新的研究方向 疫苗。