Laboratory And Preclinical Studies Of Flaviviruses

黄病毒的实验室和临床前研究

• 批准号: 10272083
• 负责人: Stephen Whitehead
• 金额: $ 105.79万
• 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/10272083
• 关键词: Accounting; Admixture; Antibodies; Asia; Attention; Attenuated; Attenuated Live Virus Vaccine; Basic Science; Binding; Biological Assay; Brain; Case Study; Cells; Cessation of life; China; Clinical; Clinical Distribution; Clinical Research; Clinical Trials; Collaborations; Consultations; Data; Dengue Fever; Dengue Vaccine; Dengue Virus; Development; E protein; Engineering; Epididymis; Epitopes; Evaluation; Family; Flaviviridae; Flavivirus; Formulation; Freeze Drying; Generations; Genome; Goals; Human; Immune; Immune response; Immunology; Individual; Infection; Innate Immune Response; Institution; Japanese Encephalitis; Japanese encephalitis virus; Laboratories; Laboratory Study; Length; Macaca mulatta; Maintenance; MicroRNAs; Modeling; Monkeys; Mus; Mutation; Pathogenicity; Pathology; Phase; Phase I Clinical Trials; Phase II Clinical Trials; Placenta; Preclinical Testing; Preparation; Process; RNA; Reagent; Recombinants; Research Personnel; Rivers; Seeds; Sequence Analysis; Serotyping; Shipping; Southeastern Asia; Sterility; Structure; Syndrome; Technology; Time; Tissues; Untranslated Regions; Vaccination; Vaccines; Viral Encephalitis; Virion; Virulent; Virus; Virus Replication; Visit; West Nile Fever; Work; Yellow Fever; ZIKA; ZIKV infection; Zika Virus; base; cGMP production; clinical application; clinical lot; human subject; immunogenicity; interest; macrophage; manufacturing process development; member; mosquito-borne; neutralizing antibody; nonhuman primate; phase 3 testing; preclinical development; preclinical study; programs; public health emergency; recombinant virus; research clinical testing; vaccine candidate; vaccine development; virology; Accounting; Admixture; Antibodies; Asia; Attention; Attenuated; Attenuated Live Virus Vaccine; Basic Science; Binding; Biological Assay; Brain; Case Study; Cells; Cessation of life; China; Clinical; Clinical Distribution; Clinical Research; Clinical Trials; Collaborations; Consultations; Data; Dengue Fever; Dengue Vaccine; Dengue Virus; Development; E protein; Engineering; Epididymis; Epitopes; Evaluation; Family; Flaviviridae; Flavivirus; Formulation; Freeze Drying; Generations; Genome; Goals; Human; Immune; Immune response; Immunology; Individual; Infection; Innate Immune Response; Institution; Japanese Encephalitis; Japanese encephalitis virus; Laboratories; Laboratory Study; Length; Macaca mulatta; Maintenance; MicroRNAs; Modeling; Monkeys; Mus; Mutation; Pathogenicity; Pathology; Phase; Phase I Clinical Trials; Phase II Clinical Trials; Placenta; Preclinical Testing; Preparation; Process; RNA; Reagent; Recombinants; Research Personnel; Rivers; Seeds; Sequence Analysis; Serotyping; Shipping; Southeastern Asia; Sterility; Structure; Syndrome; Technology; Time; Tissues; Untranslated Regions; Vaccination; Vaccines; Viral Encephalitis; Virion; Virulent; Virus; Virus Replication; Visit; West Nile Fever; Work; Yellow Fever; ZIKA; ZIKV infection; Zika Virus; base; cGMP production; clinical application; clinical lot; human subject; immunogenicity; interest; macrophage; manufacturing process development; member; mosquito-borne; neutralizing antibody; nonhuman primate; phase 3 testing; preclinical development; preclinical study; programs; public health emergency; recombinant virus; research clinical testing; vaccine candidate; vaccine development; virology

The mosquito-borne members of the Flaviviridae family, contain a single-stranded positive-sense RNA genome and are the cause of yellow fever, dengue fever, Japanese encephalitis, Zika, and West Nile fever syndromes. In recent years, much of our laboratory effort was focused on the development and preclinical testing of dengue virus vaccine candidates suitable for inclusion in a live attenuated tetravalent vaccine. Clinical lots of each of these vaccine candidates were manufactured in prior years and have been evaluated individually and in combination in numerous Phase I and II clinical trials, Optimal tetravalent admixtures have been selected and have now entered Phase III evaluation. Although the dengue virus vaccine program is predominantly in a clinical mode at this time, considerable effort is currently devoted to support a number of important functions, including, 1) manufacture, replacement, maintenance, stability/sterility analysis, and distribution of clinical lots of vaccines suitable for study in human subjects, 2) basic research on virus stabilization and lyophilization processes, 3) submission and laboratory support of IND applications for the clinical evaluation of tetravalent dengue vaccine formulations, 4) support of the seven companies/institutions that have licensed our vaccine technology or virus products, which includes consultative visits and clinical trial planning, development of manufacturing processes, preparation and shipping of vaccine seed or clinical lot viruses, assistance with sequence analysis, and sharing of IND/clinical trial data, 5) support of collaborations with investigators interested in basic virology or immunology studies, 6) use of immune cells collected from our clinical studies to investigate the innate immune response to vaccination, 7) characterization of epitopes recognized following dengue or Zika virus infection, and 8) refinement/qualification of laboratory assays such as the plaque reduction neutralization assay, and 9) isolation and recombinant construction of a suitable DENV-4 strain for use as a potential clinical challenge strain. With the emergence of Zika virus in the Western hemisphere and the recent public health emergency, our attention remains focused on the development of live attenuated vaccine candidates that would be compatible for co-formulation with our tetravalent dengue vaccine. Initially, recombinant chimeric viruses expressing the prM and E proteins of Zika virus on either the DENV-2 or DENV-4 background were generated and evaluated in rhesus monkeys and human subjects. Although the chimeric candidate appeared to be safe for use in humans, it was observed to be over-attenuated with low infectivity and subsequently limited immunogenicity. Additional candidates are currently undergoing preclinical development, in collaboration with LID, and include full-length ZIKV candidates containing 3'-UTR deletions and microRNA targets that restrict virus replication in tissues such as brain, placenta, epididymis, and certain macrophage types. Three of these candidates have been sent to Charles River Laboratories for cGMP production. In addition, collaborative studies continue to look at the pathology and immune responses of ZIKV in monkey models of infection. Mosquito-borne Japanese encephalitis virus (JEV) causes the most important viral encephalitis in the Asia Pacific region, accounting for more than 20,000 reported cases and 6,000 deaths annually. Efforts to develop a JEV vaccine continue in our laboratory and it is envisioned that a suitable live attenuated JEV vaccine could be combined with our live attenuated DEN virus vaccine to create a second-generation pentavalent vaccine for the control of these viruses in Southeast Asia. The laboratory has recovered numerous engineered viruses and has evaluated their pathogenicity in mice. The recombinant rJEV virus remains fully virulent in mice and provides a background for the evaluation of attenuating mutations. Sets of mutations derived from the attenuated SA14-14-2 vaccine virus produced in China have been introduced into the rJEV virus clone in order to evaluate the attenuating potential of mutations found in both the structural and non-structural genes. Following earlier evaluation in mice, a number of these recombinant viruses look suitable for evaluation in non-human primates and consideration as vaccine candidates. In addition, we have developed new reagents (recombinant virus particles) for use in a flow-cytometric-based neutralizing antibody assay.

Flaviviridae家族的蚊子传播成员包含一个单链的阳性RNA基因组，是黄热病，登革热，日本脑炎，Zika和West Nile Fever综合征的原因。近年来，我们的许多实验室努力都集中在登革热病毒疫苗候选物的开发和临床前测试，适合纳入活衰减的四载疫苗。这些候选疫苗的临床批次是在前几年生产的，并且已经在许多I期和II期临床试验中进行了单独评估，并且已经选择了最佳的四位化助剂，现在已经进入了III期评估。尽管目前，登革热病毒疫苗计划主要是在临床模式下，但目前正在努力支持许多重要功能，包括1），包括1），制造，替代，维护，维持，稳定性/不育分析，稳定性/不育分析，临床疫苗的分布，许多适用于人类研究的临床研究，以进行人类稳定及其稳定过程的研究，2）），2）2）3）属于型号的生产，3）3）3）3）。 4）支持我们的疫苗技术或病毒产品许可的七家公司/机构的支持从我们的临床研究中收集的免疫细胞研究对疫苗接种的先天免疫反应，7）表征登革热或寨卡病毒感染后识别的表位的表征，以及8）实验室测定的细化/资格，例如，斑块还原中性测定法，以及9）隔离和重组构建适合DENV-4临床的临床造成的临床造成的临床构造。 随着寨卡病毒在西半球的出现以及最近的公共卫生紧急情况，我们的注意力仍然集中在生存衰减的疫苗候选物的发展上，这些疫苗候选物将与我们的四维龙登革热疫苗共同成型兼容。最初，在恒河猴和人类受试者中生成并评估了在DENV-2或DENV-4背景上表达Zika病毒PRM和E蛋白的重组嵌合病毒。 尽管嵌合候选者似乎可以安全地用于人类，但观察到它的感染率低，随后受到限制的免疫原性。 目前，其他候选人正在与LID合作进行临床前开发，并包括包含3'-UTR缺失的全长ZIKV候选者和MicroRNA靶标，这些靶标限制了诸如大脑，胎盘，附子症和某些巨噬细胞类型的组织中病毒复制的限制。其中三名候选人已被派往查尔斯河实验室进行CGMP生产。 此外，协作研究继续研究猴子感染模型中ZIKV的病理和免疫反应。 蚊子传播的日本脑炎病毒（JEV）在亚太地区引起最重要的病毒脑炎，每年占20,000多例病例和6,000例死亡。在我们的实验室中，仍在继续开发JEV疫苗的努力，可以预见，可以将合适的活死的JEV疫苗与我们的活衰减的Den病毒疫苗结合使用，以创建第二代五角星疫苗，以控制东南亚这些病毒。该实验室已经回收了许多工程病毒，并评估了其在小鼠中的致病性。重组RJEV病毒在小鼠中仍然充满活力，并提供了评估衰减突变的背景。为了评估在结构和非结构性基因中发现的突变的衰减潜力，已将在中国产生的衰减的SA14-14-2疫苗病毒中得出的突变集已引入中。在较早的小鼠评估之后，许多这些重组病毒看起来适合于非人类灵长类动物的评估，并将其视为候选疫苗。 此外，我们开发了用于基于流量仪的中和抗体测定法中的新试剂（重组病毒颗粒）。