Monoclonal Antibody to Combat Pseudomonas Aeruginosa

对抗铜绿假单胞菌的单克隆抗体

• 批准号: 10674274
• 负责人: Joanna B Goldberg
• 金额: $ 102.18万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-15 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10674274
• 关键词: Acute; Affinity; Antibiotic Resistance; Antibodies; Antibody Therapy; Antibody titer measurement; Antigens; B-Lymphocytes; Bacteremia; Bacteria; Bacterial Antibiotic Resistance; Binding; Biological Assay; Blood; COVID-19 patient; Capsicum; Cells; Cellular Structures; Chronic; Clinical Research; Cloning; Communicable Diseases; Development; Epitope Mapping; Epitopes; Flow Cytometry; Functional disorder; Future; Goals; Grant; Hospitals; Human; Immune response; Immune system; Immunize; Immunocompromised Host; In Vitro; Individual; Industrialization; Infection; Injury; Investigation; Island; Life; Lipopolysaccharides; Lung infections; Membrane Proteins; Methods; Microfluidics; Modeling; Monoclonal Antibodies; Mus; Nature; Nosocomial Infections; Nosocomial pneumonia; Patients; Peptide Elongation Factor Tu; Peripheral Blood Mononuclear Cell; Persons; Phagocytes; Pharmacologic Substance; Physiological; Pneumonia; Polysaccharides; Process; Protein Biosynthesis; Proteins; Protocols documentation; Pseudomonas aeruginosa; Pseudomonas aeruginosa infection; Pseudomonas aeruginosa pneumonia; Reagent; Resistance profile; Role; Serotyping; Sorting; Spain; Spleen; Surface; Testing; Time; Universities; Vaccination; Vaccines; Washington; Work; antimicrobial; combat; cystic fibrosis infection; cystic fibrosis patients; fighting; fungus; hospital care; immune system function; immunogenic; immunological status; improved; innovation; interest; mouse model; novel; novel strategies; novel therapeutics; opportunistic pathogen; passive antibodies; pathogen; patient population; pneumonia model; post SARS-CoV-2 infection; response; superinfection; vaccine access

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.

项目总结/摘要。 铜绿假单胞菌是一种革兰氏阴性的条件致病菌，感染免疫功能低下的 个人，尤其是在医院。这种细菌是虚弱人群中的一种重要病原体 免疫系统、损伤和其他潜在的生理功能障碍。铜绿假单胞菌负责高达 20%的医院获得性肺炎。它是医院感染的主要原因之一， 是COVID-19患者共同感染或导致以下双重感染的最常见细菌之一 COVID-19感染尽管抗菌治疗和医院护理有所改善，但铜绿假单胞菌 在大约30%的病例中，菌血症和肺炎仍然是致命的。铜绿假单胞菌也是导致 囊性纤维化患者的慢性危及生命的肺部感染。这种细菌具有天然的抗生素抗性 并且感染一旦确定就很难治疗，没有可用的疫苗。 我们建议使用丰富的和必需的蛋白质，延伸因子-Tu（EF-Tu），作为抗体靶点的P。 铜绿。虽然它在蛋白质合成中的作用最为人所知，但我们小组和其他人的工作表明EF- Tu可以在铜绿假单胞菌上表面暴露。我们的初步结果表明，铜绿假单胞菌EF-Tu是 在急性铜绿假单胞菌肺炎小鼠模型中具有免疫原性和保护性。我们已经生成 在该模型中，EF-Tu的小鼠单克隆抗体促进铜绿假单胞菌的部分清除。 这是一个“开发新型治疗药物以对抗选定抗生素耐药性细菌的伙伴关系”。 和真菌”（RFA-AI-22-028），并代表了埃默里大学的乔安娜·戈德堡博士 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）， 博士来自华盛顿州西雅图的华盛顿大学的Marion Pepper和来自华盛顿大学的塞巴斯蒂安Alberti博士 位于西班牙帕尔马的巴利阿里群岛。使用来自小鼠的脾脏，我们将使用方案用EF-Tu免疫小鼠 优化用于保护和来自具有高滴度EF-Tu抗体的人的血液，我们将获得B- 将使用流式细胞术和纳米培养微流体阵列在单细胞水平筛选的细胞， 分别鉴定铜绿假单胞菌特异性单克隆抗体EF-Tu，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克隆抗体的疗效， 作为未来临床研究的第一步。我们认为这里提出的研究 代表了开发可用于P的新被动试剂的适当的第一步。 无论感染菌株的性质和相关的耐药性如何， 以及患者的免疫状态。通过这个项目的完成，我们将产生和 验证了10种鼠源性和10种人源性EF-Tu单克隆抗体。与Aridis合作 该项目的药物将使我们专注于开发这些用于人类的试剂的目标。