Novel DNA-Launched Attenuated Vaccine for VEE Virus

新型 DNA 启动的 VEE 病毒减毒疫苗

• 批准号: 8123876
• 负责人: Peter M. Pushko
• 金额: $ 30万
• 依托单位: MEDIGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8123876
• 关键词: 3' Untranslated Regions; Adjuvant; Adverse effects; Alphavirus; Amino Acids; Attenuated; Attenuated Live Virus Vaccine; Attenuated Vaccines; Bacteria; Biological; Biomedical Research; Bioterrorism; Categories; Cell Culture Techniques; Central America; Chinese Hamster Ovary Cell; Codon Nucleotides; Cold Chains; Collaborations; Combined Vaccines; Communicable Diseases; Culture Media; DNA; DNA Sequence Rearrangement; DNA Vaccines; Development; Disease Outbreaks; Dose; Epidemic; Epitopes; Evaluation; Flavivirus; Flushield; Foundations; Future; Genetic; Goals; Harvest; Health Personnel; Human; Human Resources; Human Virology; Hybrids; Immune response; Immunity; Immunization; In Vitro; Inbred BALB C Mice; Individual; Infection; Injection of therapeutic agent; Institutes; Investigational Drugs; Laboratories; Left; Legal patent; Length; Licensing; Life; Maryland; Medical Research; Methods; Molecular Cloning; Mus; Mutagenesis; Mutation; National Institute of Allergy and Infectious Disease; Phenotype; Population; Process; Production; Proteins; Protocols documentation; Recombinants; Recording of previous events; Research; Research Institute; Research Personnel; Risk; Safety; Secure; Signal Pathway; Solutions; Structural Genes; Subunit Vaccines; Technology; Texas; Transportation; United States; United States National Institutes of Health; Universities; Vaccinated; Vaccination; Vaccine Design; Vaccines; Venezuelan Equine Encephalitis Virus; Vertebral column; Viral Antigens; Virulent; Virus; Virus Diseases; attenuation; base; biodefense; biosafety level 3 facility; chemokine; cis acting element; cost; cytokine; design; epizootic; experience; immunogenic; immunogenicity; improved; in vivo; innovation; manufacturing process; meetings; mouse model; neutralizing antibody; new technology; nonhuman primate; novel; novel vaccines; pathogen; plasmid DNA; prevent; promoter; protein expression; prototype; research study; vaccine delivery; vaccine development; vaccine efficacy; weapons

DESCRIPTION (provided by applicant): Venezuelan Equine Encephalitis virus (VEEV) is a dangerous, NIH/NIAID category B human pathogen and a potential bioterrorism threat. Outbreaks of VEEV occur in Central America and have previously spread into the United States. The potentially devastating effects of the virus reemergence in the U.S. demand an effective vaccine to protect population. Currently, live attenuated TC-83 vaccine is used under IND protocol for vaccination of personnel at risk. The vaccine causes adverse effects, whereas some individuals do not develop neutralizing antibodies. The efforts to develop new VEEV vaccines are underway. However, because vaccine development is a lengthy process and the supply of TC-83 vaccine is limited, the U.S. may soon experience a shortage of VEEV vaccine, which can leave the U.S. population unprotected. We propose a revolutionary new technology for vaccination against VEEV and potentially, other viral diseases. Medigen has recently developed the "infectious DNA" (i-DNA) vaccination technology, which represents a unique combination of conventional DNA immunization with high efficacy of live attenuated vaccines. The key feature of this technology is that live attenuated virus is launched in vivo from i-DNA plasmid carrying a molecular clone of VEEV vaccine with enhanced safety and immunogenic features. In preliminary studies we have shown that injection in vivo of the prototype i-DNA derived from the TC-83 vaccine has successfully launched live attenuated vaccine in mice. Here we propose the development and feasibility evaluation of novel i-DNA VEEV vaccine for safe and efficient vaccination against VEEV based on the rational design of i-DNA clones and i-DNA immunization technology. In Specific Aim I, we will apply innovative "silent mutagenesis" method to introduce additional genetic changes into the prototype TC-83 i-DNA in order to secure attenuated phenotype and to generate i-DNA clones with high levels of safety and immunogenicity. The i-DNA clones will be transfected in CHO cells and live attenuated viruses will be harvested from culture media. Safety of i-DNA-derived viruses will be evaluated in mice in collaboration with the Institute of Human Virology, University of Maryland. In Specific Aim II we will vaccinate BALB/c mice with i-DNA constructs in order to induce immune response and to evaluate immunogenicity in vivo. Further, the efficacy of the most promising i-DNA VEEV vaccine will be evaluated in a virulent VEEV challenge experiment at Southwest Foundation for Biomedical Research (SFBR), San Antonio, TX. Finally, reversion studies in vivo will be also conducted at SFBR to demonstrate genetic stability of i-DNA - derived VEEV vaccine. The goal of this 2-year research is the identification of i- DNA VEEV vaccine clone with the optimal safety and immunogenicity profiles for future evaluation and challenge experiments in non-human primates. Our preliminary results suggest that the rational vaccine design and i-DNA technology can provide a revolutionary solution for VEEV vaccine by improving safety, genetic stability, and immunogenicity, and by eliminating many costly steps of the conventional manufacturing process. Essentially, live attenuated vaccine will be "manufactured" within the immunized individuals. This technology also utilizes many advantages of DNA vaccines (genetic homogeneity and stability, low cost of manufacturing, storage, and transportation, no cold chain) and, more importantly, enhances immunogenicity. Recombinant i-DNA clones produced in bacteria contains CpG motifs that activate TLR9 and MyD88-dependent signaling pathways resulting in robust production of cyto- and chemokines, which induce strong priming effects and stimulate acquired virus- specific immune responses. The final i-DNA VEEV vaccine will represent a novel class of vaccines combining the advantages of DNA and live attenuated vaccines and preserving the backbone of a "classic" vaccine with a history of human use. The i-DNA technology can be easily adapted for the development of other vaccines including live attenuated vaccines for WEEV, EEEV, other alphaviruses, and flaviviruses. If successful, this technology can potentially transform the field of live attenuated vaccines for many viral diseases. PUBLIC HEALTH RELEVANCE: In this application, we propose the development of a revolutionary new technology for vaccination against VEEV by using the "infectious DNA" (i-DNA), a unique hybrid of DNA and live attenuated vaccines. The key feature of this technology is that live attenuated vaccine virus is launched in vivo from i-DNA plasmid. In this study, we will generate several i-DNA vaccines and evaluate their safety, immunogenicity, and efficacy against virulent VEEV challenge in experimental mouse models. If successful, this technology can potentially transform the field of live attenuated vaccines for many viral diseases.

描述（由申请人提供）：委内瑞拉马脑炎病毒（VEEV）是一种危险的，NIH/NIAID类别B人类病原体和潜在的生物恐怖主义威胁。 VEEV的爆发发生在中美洲，以前已经蔓延到美国。该病毒再次出现在美国的潜在毁灭性影响要求有效的疫苗保护人群。目前，根据IND方案使用了活衰减的TC-83疫苗，以疫苗接种处于危险中的人员。该疫苗会引起不良反应，而某些人则不会产生中和抗体。开发新VEEV疫苗的努力正在进行中。但是，由于疫苗开发是一个漫长的过程，而TC-83疫苗的供应有限，因此美国可能很快遭受VEEV疫苗的短缺，这可能会使美国人口不受保护。 我们提出了一种革命性的新技术，用于疫苗接种VEEV以及其他病毒疾病。 Medigen最近开发了“感染性DNA”（I-DNA）疫苗接种技术，该技术代表了常规DNA免疫和活疫苗的高疗效的独特组合。该技术的关键特征是，活体病毒是由I-DNA质粒在体内发射的，该病毒含有增强的安全性和免疫原性特征的VEEV疫苗的分子克隆。在初步研究中，我们表明，从TC-83疫苗衍生的原型I-DNA的体内注射成功地发射了小鼠的活疫苗。在这里，我们提出了新型I-DNA VEEV疫苗的开发和可行性评估，以根据I-DNA克隆和I-DNA免疫技术的合理设计对VEEV进行安全有效的疫苗接种。 在特定的目标I中，我们将应用创新的“无声诱变”方法，将其他遗传变化引入原型TC-83 I-DNA，以确保衰减表型并产生具有高度安全性和免疫原性的I-DNA克隆。 I-DNA克隆将在CHO细胞中转染，并将从培养基中收集活病毒。 I-DNA衍生的病毒的安全性将在小鼠中与马里兰大学人类病毒学研究所合作评估。在特定的目标II中，我们将用I-DNA构建体接种BALB/C小鼠，以诱导免疫反应并评估体内免疫原性。此外，最有希望的I-DNA VEEV疫苗的功效将在德克萨斯州圣安东尼奥市西南生物医学研究基金会（SFBR）的一项有毒的VEEV挑战实验中进行评估。最后，还将在SFBR上进行体内的重新研究，以证明I -DNA衍生的VEEV疫苗的遗传稳定性。这项为期两年的研究的目的是鉴定具有最佳安全性和免疫原性特征的I-DNA VEEV疫苗克隆，以进行未来的评估和挑战非人类灵长类动物的挑战实验。 我们的初步结果表明，理性疫苗设计和I-DNA技术可以通过提高安全性，遗传稳定性和免疫原性，并消除常规制造过程中许多昂贵的步骤，从而为VEEV疫苗提供革命性的解决方案。本质上，在免疫个体内将“生产”活疫苗“生产”。该技术还利用了DNA疫苗（遗传均匀性和稳定性，低成本，存储和运输的低成本，没有冷链）的许多优势，更重要的是，提高了免疫原性。在细菌中产生的重组I-DNA克隆含有CPG基序，可激活TLR9和MYD88依赖性信号传导途径，从而导致稳健产生细胞环和趋化因子的产生，从而诱导强大的启动效应并刺激获得性的病毒特异性免疫反应。 最终的I-DNA VEEV疫苗将代表一种新型的疫苗，结合了DNA和活衰减的疫苗的优势，并保留具有人类使用史的“经典”疫苗的骨架。 I-DNA技术可以很容易地适应其他疫苗的开发，包括WEEV，EEEV，其他α病毒和黄病毒的活疫苗。如果成功，该技术可能会改变许多病毒疾病的活疫苗领域。 公共卫生相关性：在此应用程序中，我们建议通过使用“感染性DNA”（i-DNA）的革命性新技术来疫苗接种VEEV，这是DNA的独特混合物和活衰减的疫苗。该技术的关键特征是通过I-DNA质粒在体内发射活疫苗病毒。在这项研究中，我们将生成几种I-DNA疫苗，并评估其在实验小鼠模型中针对有毒VEEV挑战的安全性，免疫原性和功效。如果成功，该技术可能会改变许多病毒疾病的活疫苗领域。