Synthetic Generation of a Chlamydia Vaccine

衣原体疫苗的合成生成

基本信息

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
项目类别：
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

项目摘要

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

项目总监/首席研究员（最后、第一、中间）：Coleman, Matthew A 项目摘要沙眼衣原体是全球细菌性传播感染 (STI) 的最常见原因，影响每年有超过 9000 万人失明，是全球可预防性失明的最常见原因。在美国，由沙眼衣原体引起的性传播感染每年造成数十亿美元的损失（Gunn 等，1998）。因为感染可以如果没有症状，可能会多年未得到治疗，并可能导致盆腔炎、宫外孕和不孕症。因此，公共卫生需要疫苗来预防沙眼衣原体引起的疾病是合理的。尽管为开发衣原体疫苗付出了巨大努力，但尚未取得进展。研究表明，免疫接种衣原体主要外膜蛋白 (MOMP) 可以在体内诱导针对感染和疾病的显着保护作用小鼠和非人类灵长类动物（如果其天然结构得以保留）。然而，MOMP 疫苗的配制是一个主要问题。给定 MOMP 的障碍有 16 个跨膜结构域，疏水性为 40%，组装为同源三聚体并包含多个半胱氨酸可以形成二硫桥。我们现在已经证明我们可以生产三聚体，SDS- 使用纳米脂蛋白颗粒 (NLP) 的 MOMP 的抗性和活性形式。这一突破是通过将合成生物学方法和 MOMP 与载脂蛋白的无细胞共表达相结合。这个提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技术使用来自鼠衣原体的 MOMP，并添加佐剂以包含在纳米颗粒中包括我们的新型疫苗。我们这项工作有两个目标：1) 设计合成鼠 MOMP 以实现高水平协同用载脂蛋白表达和纯化以形成纳米圆盘复合物，以及 2) 在小鼠中显示出针对使用工程化、佐剂的 MOMP-NLP 颗粒进行鼻内攻击。页面延续格式页面