Amyloid-inspired Vaccine Delivery for the Elderly

为老年人提供淀粉样蛋白疫苗

• 批准号: 10457013
• 负责人: Meredith E. Jackrel
• 金额: $ 19.69万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10457013
• 关键词: 2019-nCoV; Adjuvant; Adopted; Adult; Affinity; Age; Aging; Agonist; Amyloid; Amyloid beta-Protein; Animal Model; Antibody Response; Antigen Presentation; Antigens; Autophagocytosis; Biochemical; Biophysics; CD8-Positive T-Lymphocytes; CD8B1 gene; Cells; Cellular Immunity; Cellular Morphology; Chronic; Clinical; Containment; Country; Degradation Pathway; Development; Disease; Dose; Elderly; Epitopes; Flow Cytometry; Fluorescence Microscopy; Goals; Herpes zoster disease; Hospitalization; Human; Immune; Immune System Diseases; Immune response; Immune system; Immunity; Immunohistochemistry; Immunologic Adjuvants; Impairment; Incidence; Infant; Infection; Inflammation; Inflammatory Response; Influenza; Influenza A virus; Injections; Lead; Length; Life Expectancy; Link; Lung; MF59; Memory; Microscopy; Modeling; Morbidity - disease rate; Mus; Natural Immunity; Nature; Neuraminidase; Neutrophil Infiltration; Outcome Study; Pathway interactions; Peptides; Pharmacology; Population; Predisposition; Proteins; Reporting; Research; Safety; Severities; Short-Term Memory; Site; Structure; Subunit Vaccines; Swelling; System; T cell response; T memory cell; T-Lymphocyte; T-Lymphocyte Epitopes; Thick; Time; Tissues; Toll-like receptors; Toxic effect; Transgenic Organisms; Translations; Vaccinated; Vaccination; Vaccines; Validation; Virus; Western Blotting; Work; Yeast Model System; Yeasts; age related; aged; alpha synuclein; base; beta pleated sheet; cytotoxic CD8 T cells; delivery vehicle; eosinophil; genome wide screen; immune function; immunogenicity; immunosenescence; improved; improved outcome; influenza infection; influenza pneumonia; influenza virus vaccine; insight; interest; long term memory; monocyte; nanofiber; nanomaterials; neutralizing antibody; overexpression; programmed cell death protein 1; receptor; respiratory pathogen; response; systemic inflammatory response; vaccination schedule; vaccine delivery; vaccine development; vaccine efficacy; vaccine evaluation; vaccine response; young adult

PROJECT SUMMARY Age-related decline in immunity and chronic inflammation significantly contribute to the incidence and susceptibility of the elderly to serious vaccine-preventable conditions including influenza, pneumonia, and herpes zoster. Immune function wanes in all adults as they age into their fifth decade and beyond. Current vaccines are not clinically effective and have to be administered with adjuvants to improve efficacy. Studies comparing vaccine responses in older and younger adults have shown that adjuvant-related inflammation is detrimental to vaccine efficacy in older adults. Therefore, platforms that can improve immune responses without overt inflammation are attractive for vaccine development for the elderly. We are interested in the development of peptide nanofibers as vaccine delivery vehicles for the elderly. These nanofibers trigger autophagy, which can induce an immune response without the use of inflammation-inducing adjuvants. In the current project, we will assess the toxicity and clearance of peptide nanofiber vaccines using yeast models and assess their immunogenicity and efficacy in aged mice using an influenza model. In aim 1, we will express peptide nanofibers in yeast with a GFP tag along with relevant controls to assess toxicity. We will vary the repeat length and expression conditions and confirm expression via immunoblotting. We will also analyze the cells using microscopy to confirm that cell morphology is not altered and assess clearance of the nanofibers over time. Also, the peptide nanofibers will be expressed in autophagy-deficient yeast and clearance of puncta over time will be followed using microscopy. We will also conduct a genome-wide screen to determine if peptide nanofibers engage and interact with other proteins in the cell that might provide insight into the mechanisms of action. In aim 2, we will assess the ability of peptide nanofiber vaccines to elicit robust cellular immunity and protect against lethal influenza infection using aged mice. Following a systemic prime and intranasal boost with the nanofiber vaccines bearing the model antigen OVA, we will investigate effector responses, short-term memory, and long- term memory using flow cytometry. Inflammatory response at the injection site will be assessed using footpad thickness and immunohistochemistry. Protection will be assessed in vaccinated mice following infection with a lethal dose of transgenic influenza A virus expressing the OVA epitope. Completion of the proposed work will leverage peptide nanofiber vaccines as an attractive platform for generating protective immune responses in the elderly with minimal inflammation.

项目概要 与年龄相关的免疫力下降和慢性炎症显着导致发病率和 老年人对严重的疫苗可预防疾病的易感性，包括流感、肺炎和疱疹 带状疱疹。所有成年人的免疫功能随着年龄的增长而减弱，尤其是在五岁及以上。目前的疫苗有 临床上效果不佳，必须配合佐剂以提高疗效。比较疫苗的研究 老年人和年轻人的反应表明，佐剂相关的炎症对疫苗有害 对老年人的功效。因此，可以改善免疫反应而不引起明显炎症的平台是 对老年人疫苗的开发具有吸引力。我们对肽纳米纤维的开发感兴趣 作为老年人的疫苗运输工具。这些纳米纤维触发自噬，从而诱导免疫 无需使用炎症诱导佐剂即可产生反应。 在当前的项目中，我们将使用酵母模型评估肽纳米纤维疫苗的毒性和清除率 并使用流感模型评估其在老年小鼠中的免疫原性和功效。在目标1中，我们将表达 酵母中带有 GFP 标签的肽纳米纤维以及用于评估毒性的相关对照。我们会改变重复次数 长度和表达条件并通过免疫印迹确认表达。我们还将使用以下方法分析细胞 显微镜检查以确认细胞形态没有改变并评估纳米纤维随时间的清除情况。还， 肽纳米纤维将在自噬缺陷的酵母中表达，并且随着时间的推移，斑点的清除将 随后使用显微镜。我们还将进行全基因组筛选，以确定肽纳米纤维是否 与细胞中的其他蛋白质接合并相互作用，这可能有助于深入了解其作用机制。在 目标 2，我们将评估肽纳米纤维疫苗引发强大的细胞免疫和预防感染的能力 使用老年小鼠进行致命的流感感染。使用纳米纤维进行全身灌注和鼻内增强后 带有模型抗原 OVA 的疫苗，我们将研究效应反应、短期记忆和长期记忆 使用流式细胞仪进行术语记忆。将使用足垫评估注射部位的炎症反应 厚度和免疫组织化学。将在感染疫苗后对接种疫苗的小鼠进行保护性评估 表达 OVA 表位的转基因甲型流感病毒的致死剂量。完成拟议的工作将 利用肽纳米纤维疫苗作为一个有吸引力的平台，在体内产生保护性免疫反应 炎症轻微的老年人。