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.

项目摘要/摘要。 铜绿假单胞菌是一种革兰氏阴性机会致病菌，可感染免疫功能低下的人 个人，尤其是在医院环境中，这种细菌是虚弱人群的重要病原体。 免疫系统、损伤和其他潜在的生理功能障碍是由铜绿假单胞菌引起的。 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，已引起人们的注意。 是同时感染 COVID-19 患者或引起重复感染的最常见细菌之一 尽管抗菌治疗和医院护理有所改善，但铜绿假单胞菌感染仍然存在。 菌血症和肺炎在约 30% 的病例中仍然是致命的，也是导致该病的主要原因。 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耐药性。 众所周知，感染一旦发生就很难治疗，而且没有疫苗。 我们建议使用丰富且必需的蛋白质延伸因子-Tu (EF-Tu) 作为 P. 虽然 EF- 因其在蛋白质合成中的作用而闻名，但我们小组和其他人的工作表明 EF- Tu 可以表面暴露于铜绿假单胞菌上。我们的初步结果表明，铜绿假单胞菌 EF-Tu 是。 我们已经建立了对小鼠的免疫原性和对急性铜绿假单胞菌肺炎小鼠模型的保护作用。 在此模型中，针对 EF-Tu 的小鼠单克隆抗体可促进铜绿假单胞菌的部分清除。 这个“开发新疗法来对抗特定抗生素耐药细菌的伙伴关系” 和真菌”（RFA-AI-22-028），代表了埃默里大学 Joanna Goldberg 博士之间的合作成果 佐治亚州亚特兰大大学（PI）、加利福尼亚州洛斯加托斯 Aridis Pharmaceuticals 的 Vu Truong 博士（工业合作伙伴）、 华盛顿大学西雅图分校的 Marion Pepper 博士和该大学的 Sebastian Alberti 博士 我们将使用来自西班牙帕尔马巴利阿里群岛的小鼠脾脏，按照方案对小鼠进行 EF-Tu 免疫。 针对具有高滴度 EF-Tu 抗体的人类的保护和血液进行优化，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上筛选细胞， 分别鉴定铜绿假单胞菌 EF-Tu 特异性单克隆抗体，然后进行功能筛选 我们还将测试这些单克隆抗体的功效。 我们相信这里提出的研究是小鼠感染模型作为未来临床研究的第一步。 开发可用于 P. 的新型被动试剂的适当的第一步。 铜绿假单胞菌感染患者，无论感染菌株的性质和相关抗生素耐药性如何 完成该项目后，我们将生成并了解患者的免疫状况。 与 Aridis 合作验证了 10 种鼠源和 10 种人源单克隆抗体。 该项目的药品将使我们专注于开发这些人类使用的试剂的目标。