Amyloid-inspired Vaccine Delivery for the Elderly

为老年人提供淀粉样蛋白疫苗

• 批准号: 10300848
• 负责人: Meredith E. Jackrel
• 金额: $ 23.63万
• 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 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10300848
• 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; Immunoblotting; 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; Work; Yeast Model System; Yeasts; age related; aged; alpha synuclein; base; beta pleated sheet; cytotoxic CD8 T cells; 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.

项目总结 与年龄相关的免疫力下降和慢性炎症显著地导致了 老年人对流感、肺炎和疱疹等严重的疫苗可预防疾病的易感性 带状疱疹。随着年龄的增长，所有成年人的免疫功能都会减弱。目前的疫苗是 临床上无效，必须与佐剂一起使用才能提高疗效。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 对老年人的疗效。因此，可以改善免疫反应而不出现明显炎症的平台是 对老年人的疫苗开发具有吸引力。我们对多肽纳米纤维的发展很感兴趣 作为老年人的疫苗运送工具。这些纳米纤维触发自噬，这可以诱导一种免疫 在不使用致炎佐剂的情况下做出反应。 在目前的项目中，我们将使用酵母模型评估多肽纳米纤维疫苗的毒性和清除性。 并使用流感模型评估它们在老龄小鼠中的免疫原性和有效性。在目标1中，我们将表示 酵母中带有GFP标签的多肽纳米纤维以及评估毒性的相关对照。我们会改变重复的 长度和表达条件，并通过免疫印迹确认表达。我们还将使用以下工具分析细胞 显微镜以确认细胞形态没有改变，并评估纳米纤维随时间的清除。另外， 多肽纳米纤维将在自噬缺陷的酵母中表达，随着时间的推移，点状结构的清除将是 随后用显微镜观察。我们还将进行全基因组筛查，以确定多肽纳米纤维 参与并与细胞中的其他蛋白质相互作用，可能有助于深入了解其作用机制。在……里面 目的2，我们将评估多肽纳米纤维疫苗诱导强大的细胞免疫和预防 使用老年小鼠进行致命的流感感染。在使用纳米纤维进行全身注射和鼻腔注射后 携带模型抗原OVA的疫苗，我们将研究效应器反应、短期记忆和长期记忆。 用流式细胞术检测术语记忆。注射部位的炎症反应将使用足垫进行评估 厚度和免疫组织化学。将在接种疫苗的小鼠中评估保护作用，这些小鼠感染了 表达OVA表位的转基因甲型流感病毒的致死量。拟议工作完成后， 利用多肽纳米纤维疫苗作为产生保护性免疫反应的有吸引力的平台 炎症轻微的老年人。