Amyloid-inspired Vaccine Delivery for the Elderly

项目摘要 与免疫力下降和慢性炎症相关的免疫力下降显著有助于发病率， 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，进入第五个十年及以后。目前的疫苗 临床上无效，并且必须与佐剂一起施用以提高功效。疫苗比较研究 老年人和年轻人的反应表明，与腹泻相关的炎症对疫苗接种不利， 老年人的功效。因此，可以在没有明显炎症的情况下改善免疫反应的平台是 对老年人的疫苗开发很有吸引力。我们对肽纳米纤维的发展很感兴趣 作为老年人的疫苗运输工具。这些纳米纤维触发自噬，可以诱导免疫系统 反应，而不使用炎症诱导佐剂。 在目前的项目中，我们将使用酵母模型评估肽疫苗的毒性和清除 并使用流感模型评估它们在老年小鼠中的免疫原性和功效。在目标1中，我们将表达 肽纳米纤维在酵母中与GFP标签沿着与相关的控制，以评估毒性。我们将改变重复 长度和表达条件，并通过免疫印迹确认表达。我们还将使用 通过显微镜检查来确认细胞形态没有改变，并评估纳米纤维随时间的清除。还有， 肽纳米纤维将在自噬缺陷酵母中表达， 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 参与并与细胞中的其他蛋白质相互作用，这些蛋白质可能提供对作用机制的深入了解。在 目的2，我们将评估肽类疫苗引发强大的细胞免疫和保护免受 致命的流感感染。在全身预充和鼻内加强后， 携带模型抗原OVA的疫苗，我们将研究效应反应，短期记忆和长期记忆。 使用流式细胞术的术语记忆。将使用脚垫评估注射部位的炎症反应 厚度和免疫组化。将在接种疫苗的小鼠中在感染一种 表达OVA表位的转基因甲型流感病毒的致死剂量。完成拟议工作将 利用肽类疫苗作为一个有吸引力的平台， 有轻微炎症的老年人

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.