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)

基本信息

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
项目类别：
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

项目摘要

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.

项目概要疫苗接种也许是我们这个时代最伟大的公共卫生成就。随着抗生素的爆发抗药性，开发针对多重耐药（MDR）细菌病原体的新疫苗更重要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，导致囊性纤维化 (CF) 患者严重感染、烧伤、严重伤口、肺炎以及需要插管或导尿的危重患者。清除 Pa 已成为问题，因为它已经抗生素耐药性越来越强。负面影响的最大风险因素加剧了这种情况与 MDR Pa 相关的结果是高龄。 60岁以后明显增加 MDR Pa 导致的发病率和死亡率。虽然有 Pa 疫苗正在开发中，但还没有获得许可。 R21 的目标是为我们的预防药物定义稳定的纳米颗粒 (NP) 悬浮液 Pa 疫苗可在生物膜建立之前预防 Pa，无论菌株如何。与许多革兰氏阴性病原体一样，PAO1/PA14 分支的 Pa 菌株具有 III 型分泌系统（T3SS）可以避免宿主先天免疫，并且是启动感染所必需的。 T3SS 装置（T3SA）在结构上类似于带有外部针头的分子注射器，提供了从细菌到宿主细胞的通电导管，用于运输介导关键的效应蛋白感染的方面。针尖蛋白和两个易位蛋白中的第一个定位于针尖的远端 T3SA 针介导宿主细胞接触。在Pa中，这些蛋白质分别是PcrV和PopB，并且它们是发病机制所必需的。它们在所有铜绿假单胞菌菌株中也高度保守 (95-98%) 拥有 T3SS。我们将 PcrV 和 PopB 融合得到 PaF。促进抗原的同时摄取和抗原呈递细胞佐剂，我们将 dmLT 的活性部分 LTA1 基因融合到 N- PaF (L-PaF) 末端。当受到 L-PaF 挑战时，L-PaF 显着降低小鼠和大鼠的肺 Pa 负荷 PAO1/PA14 进化枝 Pa。最近，已鉴定出 PAO7 进化枝的 Pa 异常值，这些异常值缺乏 T3SS 而是使用外溶素 A (ExlA) 来破坏宿主细胞膜。我们已将 ExlA 添加到 L-PaF 中配方，当鼻内给药时，已证明对 PAO1/14 和 PAO7 具有保护作用小鼠中的进化枝。这些小鼠的血清表现出显着的调理吞噬杀伤作用 (OPK)。此外，升高 L-PaF 疫苗接种小鼠的肺细胞分泌 IL-17 水平。 IL-17 和 OPK 均被视为对于清除 Pa 感染很重要。在此 R21 中，我们将评估稳定颗粒 ExlA/L-PaF NP 的保护性免疫反应暂停。我们将完成这个项目的两个目标：1）我们将为 ExlA/L-PaF 抗原并评估其稳定性。 2) 然后我们将确定由以下引起的免疫反应 ExlA/L-PaF 纳米颗粒制剂并确定其从肺部清除 Pa 的能力。