Synthetic Generation of a Chlamydia Vaccine

项目主任/首席调查员(最后、第一、中间)：Coleman，Matthew A 项目摘要 沙眼衣原体是世界范围内细菌性传播感染(STI)的最常见原因，影响 每年超过9000万人，是全世界最常见的可预防失明的原因。在美国， 沙眼衣原体引起的性传播感染每年造成数十亿美元的成本(Gunn等人。1998年)。因为感染可以 无症状，可能会多年得不到治疗，并可能导致盆腔炎、宫外孕和 不孕不育。因此，公共卫生需要一种疫苗来预防沙眼衣原体引起的疾病是合理的。尽管 开发衣原体疫苗的相当大的努力还没有到来。研究表明，免疫接种 衣原体主要外膜蛋白(MOMP)可诱导对感染和疾病的显著保护作用 老鼠和非人类灵长类动物，如果它的天然结构被保存下来的话。然而，MOMP疫苗的研制是一个主要的 给予MOMP的障碍有16个跨膜结构域，40%疏水，以同源三聚体的形式组装，含有 可以形成二硫键的多个半胱氨酸。我们现在已经证明，我们可以生产一种三聚体，十二烷基硫酸钠- 使用纳米脂蛋白颗粒(NLP)的抗药性和活性形式的MOMP。这一突破是通过以下方式实现的 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 重点是进一步开发以MOMP为疫苗平台的NLP。我们将重点演示这一点 使用来自衣原体的MOMP技术，将佐剂设计为包含在纳米颗粒中，以 构成了我们的新型疫苗。我们的这项工作有两个目标：1)工程合成小鼠MOMP，用于高水平的联合 用载脂蛋白表达和纯化形成纳米盘复合体，以及2)显示对小鼠的保护作用 鼻腔内挑战使用工程的，佐剂，MOMP-NLP颗粒。 页面续订格式页面

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