Synthetic Generation of a Chlamydia Vaccine

衣原体疫苗的合成生成

• 批准号: 9307728
• 负责人: Matthew Adrian Coleman
• 金额: $ 21.92万
• 依托单位: LAWRENCE LIVERMORE NATIONAL SECURITY, LLC
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9307728
• 关键词: Adjuvant; Affect; Alpha Cell; Antibiotic Therapy; Antibodies; Antibody titer measurement; Antigens; Apolipoproteins; Bacteria; Bacteriorhodopsins; Biological Assay; Biomedical Engineering; Blindness; Body Weight; Cells; Chlamydia; Chlamydia muridarum; Chlamydia trachomatis; Codon Nucleotides; Complex; Cysteine; Development; Disease; Disulfides; Ectopic Pregnancy; Engineering; Environment; Epitopes; Evaluation; Formulation; Future; Generations; Goals; Hydrogenase; Hydrophobicity; Immune response; Immunization; Immunize; Immunoglobulin A; Immunoglobulin G; In Vitro; Inbred BALB C Mice; Infection; Infertility; Interferon Type II; Intramuscular; Label; Lipids; Lung; Measures; Membrane Proteins; Modeling; Mus; Mutagenesis; Pelvic Inflammatory Disease; Pregnancy; Principal Investigator; Production; Protein Conformation; Proteins; Protocols documentation; Public Health; Reaction; Recombinants; Reproductive system; Research; Resistance; Route; Serum; Sexually Transmitted Diseases; Solubility; Structure; Synthetic Genes; Techniques; Technology; Testing; Translations; Transmembrane Domain; United States; Uterus; Vaccinated; Vaccination; Vaccines; Vagina; Variant; Weight; Woman; Work; base; cell mediated immune response; chronic abdominal pain; cost; experimental study; in vivo; major outer membrane protein; nanodisk; nanoparticle; nonhuman primate; novel; novel vaccines; particle; pathogen; prevent; programs; protein complex; protein expression; protein folding; response; scaffold; scale up; synthetic biology; vaccine candidate; vaccine delivery; vaccine development

Program Director/Principal Investigator (Last, First, Middle): Coleman, Matthew A Project Abstract Chlamydia trachomatis is the most common cause of bacterial sexually transmitted infections (STIs) worldwide affecting over 90 million people every year and the most common cause of preventable blindness worldwide. In the United States, STIs caused by C. trachomatis account for billions of dollars in annual costs (Gunn et al. 1998). Because the infection can be asymptomatic, it may go untreated for years and can result in pelvic inflammatory disease, ectopic pregnancy, and infertility. Therefore, a public health need for a vaccine to prevent diseases caused by C. trachomatis is justified. Despite considerable efforts to develop a chlamydial vaccine none has been forthcoming. Studies have shown immunization with the Chlamydia major outer membrane protein (MOMP) can induce significant protection against infection and disease in mice and non-human primates if its native structure is preserved. However, formulation of MOMP vaccines is a major hurdle given MOMP has 16 transmembrane domains, is 40% hydrophobic, assembles as a homotrimer and contains multiple cysteine's that can form disulfide bridges. We have now demonstrated that we can produce a trimeric, SDS- resistant, and active form of MOMP using nanolipoprotein particles (NLPs). This breakthrough was achieved by combining synthetic biology approaches and cell-free co-expression of MOMP with apolipoproteins. This proposal is focused on further developing NLPs formulated with MOMP as a vaccine platform. We will focus on demonstrating this technology using MOMP from Chlamydia muridarum engineered with adjuvants for inclusion within the nanoparticle to comprise our novel vaccine. We have two aims for this work: 1) Engineer synthetic murine MOMP for high-level co- expression and purification with Apolipoproteins to form a nanodisc complex, and 2) Show protection in mice against an intranasal challenge using the engineered, adjuvanted, MOMP-NLP particles. Page Continuation Format Page

项目负责人/主要研究者（最后一位、第一位、中间一位）：科尔曼、马修A 项目摘要 沙眼衣原体是细菌性性传播感染（STI）的最常见原因， 每年有超过9000万人失明，也是全球可预防失明的最常见原因。在美国， 由C.沙眼每年花费数十亿美元（古恩et al.1998）。因为感染可以 如果没有症状，可能会多年不治疗，并可能导致盆腔炎、宫外孕， 不孕因此，公共卫生需要一种疫苗来预防由C.沙眼是合理的。尽管 为研制衣原体疫苗作出了相当大的努力，但没有任何进展。研究表明， 主要衣原体外膜蛋白（MOMP）可以对感染和疾病产生显着的保护作用， 小鼠和非人类灵长类动物，如果其天然结构被保留。然而，MOMP疫苗的配制是一个主要的问题。 提供的障碍MOMP具有16个跨膜结构域，是40%疏水性的，组装为同源三聚体，并含有 可以形成二硫桥的多个半胱氨酸。我们现在已经证明，我们可以产生一个三聚体，SDS- 使用纳米脂蛋白颗粒（NLP）的MOMP的抗性和活性形式。这一突破是由 结合合成生物学方法和MOMP与载脂蛋白的无细胞共表达。这项建议是 专注于进一步开发以MOMP作为疫苗平台配制的NLP。我们将重点展示这一点 使用来自小鼠衣原体的MOMP的技术，所述MOMP用佐剂工程化以包含在纳米颗粒内， 组成了我们的新型疫苗我们的工作有两个目标：1）设计合成小鼠MOMP，用于高水平的co- 用载脂蛋白表达和纯化以形成纳米盘复合物，和2）在小鼠中显示针对免疫缺陷病毒的保护作用。 使用工程化的佐剂化的MOMP-NLP颗粒进行鼻内攻击。 页面延续格式页面