Monoclonal Antibody to Combat Pseudomonas Aeruginosa

项目摘要/摘要。 铜绿假单胞菌是一种革兰氏阴性的机会致病菌，感染免疫功能低下的 个人，特别是在医院环境中。这种细菌是虚弱的人的重要病原体 免疫系统、损伤和其他潜在的生理功能障碍。铜绿假单胞菌对高达 占所有医院获得性肺炎的20%。它是医院感染的主要原因之一，并已被注意到 是新冠肺炎合并感染患者或以下患者引起双重感染的最常见细菌之一 新冠肺炎感染。尽管在抗菌治疗和医院护理方面有所改善，但铜绿假单胞菌 菌血症和肺炎在大约30%的病例中仍然是致命的。铜绿假单胞菌也是引起 囊性纤维化患者中威胁生命的慢性肺部感染。这种细菌对抗生素具有天然抗药性。 而且众所周知，感染一旦确诊就很难治疗，因为没有疫苗可用。 我们建议使用丰富和必要的蛋白质，延伸因子-Tu(EF-Tu)作为抗P. 铜绿假单胞菌。虽然最著名的是它在蛋白质合成中的作用，但我们团队和其他人的研究表明，EF- 铜绿假单胞菌表面可暴露TU。初步结果表明，铜绿假单胞菌Ef-Tu是 对小鼠的免疫原性和对急性铜绿假单胞菌肺炎小鼠的保护作用。我们已经产生了 在该模型中，抗EF-Tu的鼠单抗可促进铜绿假单胞菌的部分清除。 这一“合作伙伴关系”旨在开发新的治疗药物以对抗某些抗生素耐药细菌 和真菌“(RFA-AI-22-028)，代表埃默里的乔安娜·戈德伯格博士之间的合作努力 佐治亚州亚特兰大大学(PI)，加利福尼亚州洛斯加托斯Aridis制药公司的Vu Truong博士(工业合作伙伴)， 华盛顿州西雅图华盛顿大学的Marion Pepper博士和该大学的Sebastian Alberti博士 位于西班牙帕尔马的巴利阿里群岛。利用小鼠的脾，我们将按程序用EF-Tu免疫小鼠 针对针对高滴度EF-Tu抗体的人类的保护和血液进行了优化，我们将获得B- 将使用流式细胞仪和纳米培养微流控阵列在单细胞水平上筛选的细胞， 分别鉴定针对铜绿假单胞菌EF-Tu的单抗，然后进行功能筛选 用于结合和吞噬杀死铜绿假单胞菌。我们还将测试这些单抗在治疗中的有效性 小鼠感染模型，作为未来临床研究的第一步。我们相信这里提出的研究 代表了开发一种新的被动试剂的适当的第一步，该试剂可以给P。 铜绿假单胞菌感染患者，无论感染菌株的性质和相关的抗生素耐药性 档案，以及病人的免疫状态。到这个项目完成时，我们将产生和 验证了10个鼠源性和10个人源性的EF-Tu单抗。与阿里迪斯合作 该项目的药物将使我们专注于开发这些供人类使用的试剂的目标。

PROJECT SUMMARY/ ABSTRACT. Pseudomonas aeruginosa is a Gram-negative, opportunistic pathogen that infects immunocompromised individuals, especially in the hospital setting. This bacterium is an important pathogen in people with weakened immune systems, injuries, and other underlying physiologic dysfunctions. P. aeruginosa is responsible for up to 20% of all hospital-acquired pneumonias. It is one of major causes of nosocomial infections and has been noted to be one of the most common bacteria co-infecting patients with COVID-19 or causing super-infections following COVID-19 infections. Despite improvements in antimicrobial therapy and hospital care, P. aeruginosa bacteremia and pneumonia remains fatal in about 30% of cases. P. aeruginosa is also the leading cause of chronic life-threatening lung infections in cystic fibrosis patients. This bacterium is naturally antibiotic resistant and infections are notoriously difficult to treat once established, with no vaccine available. We propose using the abundant and essential protein, elongation factor-Tu (EF-Tu), as an antibody target for P. aeruginosa. While best known for its role in protein synthesis, work from our group and others indicate that EF- Tu can be surface-exposed on P. aeruginosa. Our PRELIMINARY RESULTS show that P. aeruginosa EF-Tu is immunogenic in mice and protective in a murine model of acute P. aeruginosa pneumonia. We have generated a mouse monoclonal antibody to EF-Tu promotes partial clearance of P. aeruginosa in this model. This “Partnerships for the Development of Novel Therapeutics to Combat Select Antibiotic Resistant Bacteria and Fungi” (RFA-AI-22-028) and represents a collaborative effort between Dr. Joanna Goldberg at Emory University in Atlanta, GA (the PI), Dr. Vu Truong at Aridis Pharmaceuticals in Los Gatos, CA (industrial partner), Dr. Marion Pepper from the University of Washington, Seattle, WA and Dr. Sebastian Alberti from the University of Balearic Islands in Palma, Spain. Using spleens from mice we will immunize mice with EF-Tu using protocols optimized for protection and blood from humans that have high titers of antibodies to EF-Tu, we will obtain B- cells that will be screened at single cell level using flow cytometry and a nanoculture microfluidic array, respectively, to identify monoclonal antibodies specific for P. aeruginosa EF-Tu, followed by functional screens for binding and phagocytic killing of P. aeruginosa. We will also test these monoclonal antibodies for efficacy in mouse models of infection as an initial step toward future clinical studies. We believe the studies proposed here represent the appropriate first steps towards developing a new passive reagent that could be given to P. aeruginosa-infected patients regardless of the nature of the infecting strain and associated antibiotic-resistance profile, as well as the immune status of the patient. By the completion of this project, we will have generated and validated 10 murine-derived and 10 human-derived monoclonal antibodies to EF-Tu. Partnering with Aridis Pharmaceuticals for this project will keep us focused on our goal of developing these reagents for human use.