A novel vaccine against multidrug-resistant gonorrhea

摘要 淋病是第二种最常见的细菌性传播感染-最常见的是 衣原体，通常与淋病混合感染。全球约有8000万新淋病病例 每年一次。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 包括不孕症、宫外孕和慢性盆腔疼痛。病原体淋病奈瑟菌(Ng) 临床上使用的几乎所有抗生素都对淋病产生了抗药性。对头孢曲松和头孢曲松的耐药性 阿奇霉素--推荐的一线治疗药物--预示着一个无法治愈的淋病时代。美国疾病控制与预防中心 已将Ng列为威胁级别为“紧急”的微生物。一种安全有效疫苗的研制 预防淋病是公共卫生的优先事项。此外，Ng和衣原体混合感染是一种频繁的事件。 即使在衣原体感染的情况下，成功的淋球菌疫苗也能预防感染。 我们鉴定了一种名为2C7的识别脂低聚糖(LOS)表位的单抗 在体内由95%的Ng表达，对小鼠的毒力至关重要。单抗2C7具有杀菌作用，并且显著 减少小鼠感染Ng的持续时间和负担。它的无处不在的表达，在毒力中的关键作用，以及 抗2C7表位抗体的杀菌特性使其成为一种有吸引力的疫苗抗原。绕过 LOS作为疫苗抗原的局限性，我们发现了一种模拟2C7表位的多肽，当 被配置为多抗原肽(MAP)，在小鼠中诱导杀菌抗体并减轻Ng感染。 由NIAID/NIH监督的产品开发计划使我们与ABL Inc.、多肽国际和 传染病研究所(IDRI)。人民民主党的目标已经实现，并取得了成果 设计一种新的四环结构(TMCP2)并确定GLA-SE为最佳佐剂基础 对小鼠免疫原性的影响。在目标1中，多肽国际公司将生产TMCP2，进行稳定性测试，并 将技术转让给cGMP制造商。在目标2(UMAS)中，小鼠和非人类灵长类动物将被 用发育级TMCP2+GLA-SE免疫以评估I)免疫原性和II)功能性 在Ng感染或单独感染Ng/衣原体感染的小鼠中产生抗体。NG与补体抑制剂结合， 人类特有的H因子(FH)和C4b结合蛋白(C4BP)，从而抑制疫苗的活性 阿布。因此，我们将在表达人FH和C4BP的新型转基因小鼠中测试疫苗效力。 在人体内模拟条件。从免疫猕猴中提纯的抗LOS抗体将用于检测 杀菌活性及其在单一Ng和Ng/衣原体混合感染小鼠中的定植能力。 ABL Inc.将获得ELISA和SBAS的资格。新西兰白猪的GLP级安全性和毒理学研究 兔子(ABL，Inc.)这对产品开发至关重要，将在目标3中执行。 研究将在目标4进行。这些目标的成功完成将为我们的候选疫苗做好准备 人文科学

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