Development of a broadly protective subunit vaccine against Pseudomonas aeruginosa

开发针对铜绿假单胞菌的广泛保护性亚单位疫苗

• 批准号: 9763456
• 负责人: Wendy L Picking
• 金额: $ 23.79万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-15 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9763456
• 关键词: Adjuvant; Adult; Antibiotic Resistance; Antibodies; Bacteria; Bordetella; Burkholderia; Burn injury; Catheterization; Cells; Cessation of life; Clinical; Cystic Fibrosis; Cytoplasm; Development; Disease; Distal; Dose; Foundations; Fruit; Future; Immune response; Immunity; Immunization; Infection; Intubation; Investigation; Life; Lung; Lung infections; Mediating; Microbiology; Modeling; Molecular; Mucous Membrane; Multi-Drug Resistance; Mus; Natural Immunity; Nature; Needles; Organism; Pathogenesis; Patients; Pneumonia; Procedures; Protein Chemistry; Protein Subunits; Proteins; Pseudomonas; Pseudomonas aeruginosa; Quality of life; Rattus; Respiratory physiology; Salmonella enterica; Shigella; Specialist; Structure; Subunit Vaccines; Syringes; Testing; Toxin; Type III Secretion System Pathway; Vaccinated; Vaccination; Vaccine Antigen; Vaccines; Virulence; Virulence Factors; Work; cell mediated immune response; cystic fibrosis patients; cytokine; enterotoxigenic Escherichia coli; experimental study; healthcare-associated infections; high reward; high risk; human pathogen; innovation; mouse model; mutant; novel; pathogen; protective efficacy; vaccinology; wound

SUMMARY Pseudomonas aeruginosa is a ubiquitous bacterium in nature and an important opportunistic human pathogen that can cause severe infections in patients with burns, severe wounds, pneumonia, and undergoing nosocomial procedures, especially those requiring intubation or catheterization. Clearing P. aeruginosa (Pa) has become progressively more difficult with growing antibiotic resistance. Of particular concern, Pa is the major cause of pulmonary infections in cystic fibrosis (CF) patients with over 70% of CF patients being colonized by adulthood. These infections decrease lung function that often reduce patient quality of life and cause early death. While there have been of Pa vaccines in development, none have been licensed. Like many Gram-negative pathogens, Pseudomonas spp. possess a type III secretion system (T3SS), a virulence factor that allows avoidance of host innate immunity and is required for the onset of infection. Structurally resembling a molecular syringe with an external needle, the T3SS apparatus (T3SA) provides an energized conduit from the bacterial cytoplasm into the host cell. 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. These proteins, PcrV and PopB, respectively, are required for Pseudomonas pathogenesis and are 95-98% conserved among Pseudomonas spp. The equivalent conserved Shigella T3SA tip and translocator proteins, IpaD and IpaB, respectively, were fused to give DBF, which we have shown as a novel subunit vaccine antigen protecting mice against challenges by homologous and heterologous Shigella spp. Similarly, highly conserved T3SA tip and translocator fusions have protected mice against Bordetella spp., Salmonella enterica and Burkholderia spp. In a preliminary mouse experiment, PaF, the PcrV/PopB fusion, was admixed with a mucosal adjuvant and delivered intranasally. In this initial preliminary experiment, 100% protection (with 44% sterilizing immunity) was seen for the PaF vaccinated mice. However, 60% of the PBS control mice also survived (although with no sterilizing immunity). Nevertheless, in this high risk, high reward R21, we will optimize the vaccine in the mouse model and to extend the findings to the accepted rat model of cystic fibrosis. Thus, because the PaF+dmLT vaccine appears to protect mice against Pa pulmonary challenge with 44% sterilizing immunity, PaF+dmLT will elicit a humoral and cell-mediated immune response that protects against most Pa strains, including those that cause disease in CF patients. The two specific aims of this investigation are: 1) Assess the respective humoral and cell-mediated immune responses elicited by PaF+dmLT and 2) Determine the protective efficacy of PaF+dmLT against Pa.

概括 铜绿假单胞菌是自然界中普遍存在的细菌，也是人类重要的机会致病菌。 病原体可引起烧伤、重伤、肺炎和接受治疗的患者严重感染 医院内手术，尤其是需要插管或导管插入术的手术。清除铜绿假单胞菌 (Pa) 有 随着抗生素耐药性的增加，变得越来越困难。特别值得关注的是，Pa是主要的 囊性纤维化 (CF) 患者肺部感染的原因，超过 70% 的 CF 患者体内有细菌定植 成年期。这些感染会降低肺功能，通常会降低患者的生活质量并导致过早死亡。 虽然有多种 Pa 疫苗正在开发中，但尚未获得许可。 像许多革兰氏阴性病原体一样，假单胞菌属。拥有III型分泌系统（T3SS）， 毒力因子，可以避免宿主先天免疫，并且是感染发生所必需的。 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 从细菌细胞质进入宿主细胞的通电导管。针尖蛋白和两个中的第一个 易位蛋白定位于 T3SA 针的远端以介导宿主细胞接触。这些蛋白质， PcrV 和 PopB 分别是假单胞菌发病机制所必需的，并且在 假单胞菌属等效的保守志贺氏菌 T3SA 尖端和易位蛋白 IpaD 和 IpaB， 分别融合产生 DBF，我们将其作为保护小鼠的新型亚单位疫苗抗原 对抗同源和异源志贺氏菌属的挑战。同样，高度保守的 T3SA 尖端和 易位蛋白融合可以保护小鼠免受博德特氏菌属、肠沙门氏菌和伯克霍尔德氏菌属的侵害。在 初步的小鼠实验将 PaF（PcrV/PopB 融合体）与粘膜佐剂混合， 鼻内递送。在这个最初的初步实验中，100% 的保护（具有 44% 的灭菌免疫力） 在 PaF 疫苗接种的小鼠中观察到。然而，60% 的 PBS 对照小鼠也存活下来（尽管没有 杀菌免疫力）。尽管如此，在这个高风险、高回报的R21中，我们将优化小鼠体内的疫苗 模型并将研究结果扩展到公认的囊性纤维化大鼠模型。 因此，因为 PaF+dmLT 疫苗似乎可以保护小鼠免受 Pa 肺部攻击 44% 灭菌免疫，PaF+dmLT 将引发体液和细胞介导的免疫反应， 可以预防大多数 Pa 菌株，包括那些导致 CF 患者患病的菌株。两个具体目标 这项研究的主要内容是：1）评估由以下因素引起的各自的体液和细胞介导的免疫反应： PaF+dmLT 和 2) 确定 PaF+dmLT 对 Pa 的保护功效。

项目成果

Effect of Two Unique Nanoparticle Formulations on the Efficacy of a Broadly Protective Vaccine Against Pseudomonas Aeruginosa.

两种独特的纳米颗粒制剂对铜绿假单胞菌广泛保护性疫苗功效的影响。

• 发表时间: 2021
• 作者: Howlader, Debaki R;Das, Sayan;Lu, Ti;Hu, Gang;Varisco, David J;Dietz, Zackary K;Walton, Sierra P;Ratnakaram, Siva Sai Kumar;Gardner, Francesca M;Ernst, Robert K;Picking, William D;Picking, Wendy L
• 通讯作者: Picking, Wendy L