Enhancing DNA vaccines using modified Cholera Toxin A1 Subunit as an adjuvant

使用改良霍乱毒素 A1 亚基作为佐剂增强 DNA 疫苗

• 批准号: 7337874
• 负责人: Timothy R Fouts
• 金额: $ 29.46万
• 依托单位: PROFECTUS BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-06-01 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7337874
• 关键词: ADP-Ribosylation Factors; Active Sites; Adjuvant; Adjuvanticity; Animals; Antibody Formation; Antigens; Aromatic Amino Acids; Binding; Biological Assay; Cell membrane; Cells; Chimera organism; Cholera Toxin; Clinical; Clinical Trials; DNA; DNA Vaccines; Dendritic Cells; Development; Dose; Drug Formulations; Enterotoxins; Evaluation; Exhibits; Face; Frequencies; Gagging; Goals; Government; HIV; Heating; Human; Immune response; In Vitro; Mediator of activation protein; Modeling; Modification; Mus; Mutation; Numbers; Persons; Pertussis Toxin; Phase; Phase II Clinical Trials; Plasmids; Primates; Proteins; Purpose; Reporter; Safety; Screening Result; Signal Transduction; Solubility; Thinking; Time; Toxin; Vaccination; Vaccine Adjuvant; Vaccines; base; cellular targeting; follow-up; immunogenicity; improved; in vivo; mutant; plasmid DNA; receptor; research clinical testing; response; size

DESCRIPTION (provided by applicant): Vaccination using plasmid DNA has the potential to provide enhanced safety and efficacy over conventional vaccines with increased stability, and accelerated product development. Unfortunately, vaccination of primates with DNA requires multiple inoculations using excessively large doses of plasmid. The A1 domains of cholera toxin (CTA1) and the related heat-labile enterotoxin (LTA1) have demonstrated particular promise as adjuvants that can enhance the immunogenicity of DNA vaccines in small animals and may provide a necessary dose sparing effect in primates. To maximally exploit this promise, however, the activity of CTA1 or LTA1 in vivo should be improved. Our objective is to identify modifications to CTA1 or LTA1 that will enhance their activity in vivo by pursuing the following aims: Aim 1: Identify mutations in the active site of either CTA1 or LTA1 that enhance enzymatic activity in vitro; Aim 2: Identify fusion constructs between CTA1 or LTA1 and human ARF6 that enhance enzymatic activity in vitro; and, Aim 3: Identify constructs from Aims 1, and 2 that optimally enhance the immunogenicity of a candidate SIMmac239-gag DNA vaccine in vivo. Candidate constructs that display a minimum of 10-fold enhanced adjuvanticity in vivo will be selected for evaluation in primate studies to be proposed in phase 2. Successful candidates in phase 2 will be incorporated into candidate HIV DNA vaccines intended for clinical evaluation.

描述（由申请人提供）：使用质粒DNA的疫苗接种有可能在稳定性提高和加速产物开发的情况下，对常规疫苗提供增强的安全性和功效。不幸的是，使用DNA的灵长类动物接种疫苗需要多次接种，使用过多剂量的质粒。霍乱毒素（CTA1）和相关的热能肠毒素（LTA1）的A1结构域已表现出特殊的希望，因为佐剂可以增强小动物中DNA疫苗的免疫原性，并可能在灵长类动物中提供必要的剂量差异。但是，为了最大程度地利用这一诺言，应改善CTA1或LTA1在体内的活性。我们的目标是确定对CTA1或LTA1的修改，以增强其活动 通过追求以下目标：目标1：在CTA1或LTA1的活性位点中确定突变，从而增强体外酶活性；目标2：确定CTA1或LTA1与人ARF6之间的融合构建体，从而增强体外酶促活性；并且，目标3：从AIMS 1和2中确定构造，从而最佳地增强了体内候选SIMMAC239-GAG DNA疫苗的免疫原性。 将在第2阶段提出的灵长类动物研究中选择至少显示10倍增强的辅助性的候选构建体。第2阶段的成功候选者将纳入旨在用于临床评估的候选HIV HIV DNA疫苗中。