An intranasal room stable vaccine formulation to prevent Pseudomonas aeruginosa (R21AI169691)

用于预防铜绿假单胞菌的鼻内室内稳定疫苗制剂 (R21AI169691)

• 批准号: 10741018
• 负责人: Wendy L Picking
• 金额: $ 22.97万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10741018
• 关键词: Achievement; Acute; Acute Pneumonia; Adjuvant; Age; Antibiotic Resistance; Antibiotics; Antibodies; Antigen-Presenting Cells; Antigens; Bacteria; Burn injury; Catheterization; Cell membrane; Cells; Critical Illness; Cytoplasm; Development; Distal; Dryness; Elderly; Emulsions; Exhibits; Explosion; Formulation; Goals; Health; IL17 gene; Immune response; Immunize; Infection; Infection prevention; Injury; Innate Immune System; Intubation; L Cells; Laboratories; Licensing; Lung; Manuscripts; Mediating; Microbial Biofilms; Molecular; Morbidity - disease rate; Mucous Membrane; Multi-Drug Resistance; Mus; Natural Immunity; Nature; Needles; Nose; Oils; Outcome; Particulate; Pathogenesis; Patients; Pneumonia; Powder dose form; Preparation; Procedures; Proteins; Pseudomonas aeruginosa; Pseudomonas aeruginosa infection; Public Health; Publishing; Pulmonary Challenge; Rattus; Risk; Risk Factors; Rodent; Suspensions; Syringes; Technology; Temperature; Time; Toxin; Type III Secretion System Pathway; Vaccinated; Vaccination; Vaccines; Water; biophysical techniques; clinically relevant; cystic fibrosis patients; cytokine; enterotoxigenic Escherichia coli; human pathogen; mortality; multidrug-resistant Pseudomonas aeruginosa; mutant; nanoparticle; novel vaccines; pathogen; pathogenic bacteria; prevent; prophylactic; resistant strain; uptake; vaccine formulation; wound

PROJECT SUMMARY Vaccination is perhaps the greatest public health achievement of our time. With an explosion of antibiotic resistance, developing new vaccines against multi-drug resistant (MDR) bacterial pathogens is more important than ever. Pseudomonas aeruginosa (Pa) is an important opportunistic human pathogen that causes severe infections in patients with cystic fibrosis (CF), burns, severe wounds, pneumonia, as well as critically ill patients who require intubation or catheterization. Clearing Pa has become problematic as it has become increasingly antibiotic resistant. This is exacerbated by the fact that the biggest risk factor for negative outcomes associated with MDR Pa is advanced age. After the age of 60, there is a significant increase in morbidity and mortality resulting from MDR Pa. While there are Pa vaccines in development, none are licensed. The goal of the R21 is to define a stable nanoparticle (NP) suspension for our prophylactic Pa vaccine that prevents Pa, regardless of strain, prior to establishment of a biofilm. Like many Gram-negative pathogens, Pa strains of the PAO1/PA14-clades possess a type III secretion system (T3SS) that allows avoidance of host innate immunity and is required for initiating infection. Structurally resembling a molecular syringe with an external needle, the T3SS apparatus (T3SA) provides an energized conduit from the bacterium into the host cell for transporting the effector proteins that mediate key aspects of infection. A needle tip protein and the first of two translocator proteins localize to the distal end of the T3SA needle to mediate host cell contact. In Pa these proteins are PcrV and PopB, respectively, and they are required for pathogenesis. They are also highly conserved (95-98%) among all P. aeruginosa strains that possess a T3SS. We have fused PcrV and PopB to give PaF. To promote simultaneous uptake of antigen and adjuvant by antigen presenting cells, we genetically fused LTA1, the active moiety of dmLT, to the N- terminus of PaF (L-PaF). L-PaF reduces mouse and rat lung Pa burden significantly when challenged with a PAO1/PA14 clade Pa. Recently, Pa outliers of the PAO7 clade have been identified that are devoid of the T3SS and instead use exolysin A (ExlA) to disrupt host cell membranes. We have added ExlA to our L-PaF formulation and, when delivered intranasally, have demonstrated protection against PAO1/14 and PAO7 clades in mice. Sera from these mice exhibit significant opsonophagocytic killing (OPK). Additionally, elevated levels of IL-17 were secreted from lung cells of L-PaF-vaccinated mice. Both IL-17 and OPK are deemed important in clearing Pa infections. In this R21, we will assess the protective immune response of a stable particulate ExlA/L-PaF NP suspension. We will complete this project in two aims: 1) We will generate nanoparticle formulations for the ExlA/L-PaF antigens and assess their stability. 2) We will then determine the immune response(s) elicited by the ExlA/L-PaF nanoparticle formulation(s) and determine their ability to clear Pa from the lungs.

项目摘要 疫苗接种也许是我们时代最大的公共健康成就。随着抗生素的爆炸 耐药性，开发针对多药耐药（MDR）细菌病原体的新疫苗更多是 比以往任何时候都重要。铜绿假单胞菌（PA）是一种重要的机会性人类病原体 引起囊性纤维化（CF），烧伤，严重伤口，肺炎以及 需要插管或导管插入的重症患者。清除PA已经成为问题 变得越来越多的抗生素耐药性。这是负面的最大风险因素，这加剧了这一事实 与MDR PA相关的结果是高龄。 60岁以后，大幅增加 MDR PA产生的发病率和死亡率。尽管有PA疫苗正在开发中，但没有一个 许可。 R21的目的是为我们的预防性定义稳定的纳米颗粒（NP）悬架 在建立生物膜之前，可以防止PA的PA疫苗，无论菌株如何。 像许多革兰氏阴性病原体一样，PAO1/PA14-Clades的PA菌株具有III型分泌物 系统（T3SS）允许避免宿主先天免疫，并且是启动感染所必需的。 T3SS设备（T3SA）在结构上类似于带有外针的分子注射器，提供了一个 从细菌进入宿主细胞的通向导管，用于转运介导键的效应蛋白 感染方面。针尖蛋白和两个转运蛋白中的第一个定位于远端 T3SA针介导宿主细胞接触。在PA中，这些蛋白质分别为PCRV和POPB，它们是 是发病机理所必需的。它们在所有铜绿假单胞菌菌株中也是高度保守的（95-98％） 拥有T3SS。我们已经融合了PCRV和POPB来提供PAF。促进同时吸收抗原 通过抗原呈现细胞辅助，我们在遗传上融合了DMLT的活性部分， PAF（L-PAF）的末端。当用A挑战时，L-PAF可显着减轻小鼠和肺部的负担 PAO1/PA14进化枝PA。最近，已经确定了PAO7进化枝的PA异层没有 T3SS并使用exolysin a（EXLA）破坏宿主细胞膜。我们已经在L-PAF中添加了EXLA 配方，当鼻内交付时，已显示出针对PAO1/14和PAO7的保护 小鼠中的进化枝。这些小鼠的血清表现出明显的调查性杀伤性（OPK）。另外，提高了 IL-17的水平是从L-PAF接种疫苗的小鼠的肺细胞中分泌的。 IL-17和OPK均被视为 清除PA感染很重要。 在此R21中，我们将评估稳定的颗粒EXLA/L-PAF NP的保护性免疫反应 暂停。我们将以两个目的完成该项目：1）我们将生成纳米颗粒配方 EXLA/L-PAF抗原并评估其稳定性。 2）然后我们将确定由 EXLA/L-PAF纳米颗粒制剂（S）并确定其从肺部清除PA的能力。