A novel vaccine against multidrug-resistant gonorrhea

摘要 淋病是第二种最常见的细菌性性传播感染-最常见的是 衣原体，通常与淋病合并感染。全球约有8000万新发淋病病例 每年。美国每年报告的淋病病例超过45万例。女性淋病的严重后遗症 包括不孕、宫外孕和慢性盆腔疼痛。淋病奈瑟氏菌（Ng），病原体 淋病，已成为耐几乎每一种抗生素在临床使用。对头孢曲松耐药， 阿奇霉素--推荐的一线治疗--预示着淋病无法治愈的时代的到来。疾控中心 已将Ng列为威胁级别为“紧急”的微生物。开发安全有效的疫苗 防治淋病是公共卫生的优先事项。此外，Ng和衣原体混合感染是常见事件 即使存在衣原体感染，成功的淋球菌疫苗也能预防感染。 我们鉴定了一种称为2C 7的单克隆抗体（mAb），其识别脂寡糖（LOS）表位 在体内由>95%的Ng表达，并且对于小鼠中的毒力至关重要。mAb 2C 7具有杀菌作用， 减少小鼠中Ng感染的持续时间和负担。它的普遍表达，在毒力中的关键作用， 抗2C 7表位的Ab的杀菌性质使其成为有吸引力的疫苗抗原。规避 由于LOS作为疫苗抗原的局限性，我们鉴定了2C 7表位的肽模拟物， 配置为多抗原肽（MAP），在小鼠中引发杀菌Ab并减弱Ng感染。 由NIAID/NIH监督的产品开发计划使我们与ABL公司合作，肽国际和 传染病研究所（IDRI）PDP的目标已经实现， 在设计一种新的四-MAP结构（TMCP 2）和鉴定GLA-SE作为最佳佐剂的基础上， 对小鼠免疫原性的影响在目标1中，肽国际将生产TMCP 2，进行稳定性测试， 将技术转让给cGMP生产商。在Aim 2（马萨诸塞大学）中，小鼠和非人类灵长类动物将被 用开发级TMCP 2加GLA-SE免疫，以评价i）免疫原性和ii）功能性， 在Ng感染的小鼠中或（单独地）在Ng/衣原体共感染的小鼠中引发Ab。Ng结合补体抑制剂， H因子（FH）和C4 b结合蛋白（C4 BP），抑制疫苗的活性 AB.因此，我们将在表达人FH和C4 BP的新型转基因小鼠中测试疫苗效力， 模拟人体条件。将测定从免疫恒河猴纯化的抗LOS Ab， 杀菌活性和减弱单一Ng和Ng/衣原体共感染小鼠中定殖的能力。 ABL Inc.将使ELISAs和SBAs合格。在新西兰白色动物中进行的GLP级安全性和毒理学研究 家兔（ABL，Inc.）这对产品开发至关重要，将在目标3中执行。IND前启用 将在目标4中进行研究。这些目标的成功完成将使我们的候选疫苗准备就绪， 人类研究

ABSTRACT Gonorrhea is the second most common bacterial sexually transmitted infection – the most common is chlamydia, which often coinfects with gonorrhea. About 80 million new cases of gonorrhea occur worldwide annually. Over 450,000 cases are reported yearly in the U.S. Serious sequelae of gonorrhea in women includes infertility, ectopic pregnancy and chronic pelvic pain. Neisseria gonorrhoeae (Ng), the causative agent of gonorrhea, has become resistant to almost every antibiotic in clinical use. Resistance to ceftriaxone and azithromycin – the recommended first-line of treatment –portends an era of untreatable gonorrhea. The CDC has listed Ng as a microorganism with a threat level of “Urgent”. Development of a safe and effective vaccine against gonorrhea is a public health priority. Furthermore, Ng and chlamydia coinfection is a frequent event and a successful gonococcal vaccine will prevent infection even when chlamydia infection is present. We identified a monoclonal antibody (mAb) called 2C7 that recognizes a lipooligosaccharide (LOS) epitope expressed by >95% of Ng in vivo and is critical for virulence in mice. mAb 2C7 is bactericidal and significantly reduces the duration and burden of Ng infection in mice. Its ubiquitous expression, key role in virulence, and the bactericidal nature of Ab against the 2C7 epitope makes it an attractive vaccine antigen. To circumvent limitations of LOS as a vaccine antigen, we identified a peptide mimic of the 2C7 epitope, which when configured as a multi-antigen peptide (MAP), elicited bactericidal Abs and attenuated Ng infection in mice. A Product Development Plan overseen by NIAID/NIH partnered us with ABL Inc., Peptides International and the Infectious Diseases Research Institute (IDRI). The objectives of the PDP have been met and has resulted in design of a novel tetra-MAP structure (TMCP2) and identification of GLA-SE as the optimal adjuvant based on immunogenicity in mice. In Aim 1, Peptides International will produce TMCP2, perform stability testing and transfer technology to the cGMP manufacturer. In Aim 2 (UMass), mice and non-human primates will be immunized with development-grade TMCP2 plus GLA-SE to evaluate i) immunogenicity and ii) functionality of elicited Ab in Ng infected or (separately) in Ng/chlamydia co-infected mice. Ng bind the complement inhibitors, factor H (FH) and C4b-binding protein (C4BP) in a human-specific manner, which dampens activity of vaccine Ab. Therefore, we will test vaccine efficacy in novel transgenic mice that express human FH and C4BP to simulate conditions in humans. Anti-LOS Ab purified from immunized Rhesus macaques will be assayed for bactericidal activity and for its ability to attenuate colonization in single Ng and Ng/chlamydia coinfected mice. ABL Inc. will qualify ELISAs and SBAs. A GLP-grade safety and toxicology study in New Zealand White Rabbits (ABL, Inc.) which is essential for product development will be performed in Aim 3. Pre-IND enabling studies will be carried out in Aim 4. Successful completion of these Aims will ready our vaccine candidate for human studies