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

计划主任/首席研究员（最后，第一，中间）：科尔曼，马修A 项目摘要沙眼衣原体是全世界细菌性传播感染（STI）的最常见原因每年有超过9000万人，这是全球可预防失明的最常见原因。在美国，由沙眼梭状疱疹引起的性传播感染占数十亿美元的年成本（Gunn等，1998）。因为感染可以无症状，它可能没有治疗多年，可能导致骨盆炎症性疾病，异位妊娠和不育。因此，公共卫生需要疫苗预防由沙眼梭状芽孢杆菌引起的疾病。尽管开发衣原体疫苗的巨大努力，没有任何事情正在进行。研究表明免疫衣原体主要外膜蛋白（MOMP）可以引起对感染和疾病的明显保护小鼠和非人类灵长类动物如果保留其天然结构。但是，MOMP疫苗的配方是主要的 MOMP给定的障碍具有16个跨膜结构域，为40％疏水，以同二聚体的形式组装并包含多种半胱氨酸可以形成二硫键。现在，我们已经证明我们可以产生三聚体SDS- 使用纳米蛋白蛋白颗粒（NLP）的MOMP的抗性和活性形式。这一突破是通过结合MOMP与载脂蛋白的合成生物学方法和无细胞共表达。该提议是专注于进一步开发以MOMP为疫苗平台的NLP。我们将专注于证明这一点使用佐剂设计的Chlamydia Muridarum的MOMP的技术，将纳米颗粒包含在内包括我们的新型疫苗。我们对这项工作有两个目标：1）工程师合成的鼠MOMP，用于高级共同用载脂蛋白表达和纯化以形成纳米盘复合物，2）在小鼠中显示保护使用工程辅助的MOMP-NLP颗粒进行鼻内挑战。页面延续格式页面