Effective Mucosal Vaccination through Engineered Outer Membrane Vesicles

通过工程外膜囊泡进行有效的粘膜疫苗接种

• 批准号: 9510636
• 负责人: DAVID A PUTNAM
• 金额: $ 19.21万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-25 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9510636
• 关键词: Animal Model; Antigens; Benchmarking; Biological Assay; CD8B1 gene; Cellular Immunity; Dendritic Cells; Engineering; Epithelium; Epitopes; Family suidae; Formulation; Goals; Green Fluorescent Proteins; Hemagglutinin; Human; Immune response; Immune system; Immunity; Influenza; Interleukin-10; Intestines; Investigation; Lamina Propria; Langerhans cell; Lipopolysaccharides; Lung; Measures; Membrane; Methods; Modeling; Mucosal Immunity; Mucous Membrane; Mus; Myelogenous; Oral; Oral cavity; Oral mucous membrane structure; Penetration; Peptides; Population; Recombinants; Regulatory T-Lymphocyte; Research; Route; Site; Subcutaneous Injections; Submucosa; T-Lymphocyte; TLR4 gene; Technology; Tissues; Topical application; Vaccine Design; Vaccine Production; Vaccines; Vesicle; Work; adaptive immune response; base; cell mediated immune response; clinically relevant; design; experience; innovation; lymph nodes; mucosal vaccination; nanosized; neutralizing antibody; pathogen; response; skills; subcutaneous; transmission process; unpublished works; vaccine delivery

The mucosa is a primary site for pathogen transmission, but injected vaccines induce poor mucosal immunity. Mucosal dendritic cell subsets in the buccal tissue can induce both immunity and tolerance to antigens. The proposed research focuses on inducing mucosal immunity by activating mucosal dendritic cells using recombinant outer membrane vesicles (rOMVs). Over the past decade, our research team has designed and developed rOMVs to induce robust Th1/Th2 immune responses to peptide antigens. In more recent and yet unpublished work, our team has discovered a way to activate and drive the maturation of DCs. Perhaps most exciting is that we quantified the penetration depth of rOMVs in the buccal mucosal tissue of the pig and show that the ~100 nm vesicles penetrate the tissue to depths consistent with the population of naïve myeloid dendritic cells (>100 µm). This rOMV-based technology, therefore, has a profile consistent with a mucosally-deliverable vaccine, specifically via buccal tissue. The objective of the research is to illustrate and establish the uniqueness of the rOMV approach as a method for buccal vaccine delivery. By combining the use of subunit epitopes with the tissue penetration of rOMVs, we will evaluate the hypothesis that the administration of these rOMVs to the buccal mucosa will induce the activation of naïve DCs and induce a strong and balanced Th1/Th2 immune response. The hypothesis will be evaluated through the completion of the following Aims. Aim 1: to quantify the immune response to a “weak” antigen to determine the efficacy and potency of the rOMVs. Our previous work used green fluorescent protein (GFP) as an effective model of a “weak” antigen that demonstrated the potency of rOMVs administered subcutaneously. We will use the same approach to quantify both the humoral and cellular responses to GFP delivered by rOMVs to the buccal mucosa of pigs (the closest animal model to the human buccal mucosa). The results will be compared to the same formulation administered subcutaneously (i.e., benchmark) in order to correlate the efficacy of the buccal route of administration to a traditional route of administration. Aim 2: to quantify the immune response to the hemagglutinin (HA) antigen of influenza to determine the protective immunity conferred by the rOMVs. In previous work we used a sequence of HA delivered by subcutaneous injection of rOMVs in mice to quantify the level of protective immunity, as measured by the hemagglutinin inhibition assay and the antibody neutralization analysis. We will use the same approach to quantify both the humoral and cellular responses to HA delivered by rOMVs to the buccal mucosa of pigs. The results will be compared to the same formulation administered subcutaneously (i.e., benchmark) in order to correlate the efficacy of the buccal route of administration to a traditional route of administration. The Cornell- based research team of Putnam (rOMV vaccine design) and Chang (swine immunity) combines the labs of two experts who collectively have the skill to create an effective, buccally administered vaccine delivery platform that is potentially applicable to a wide range of mucosally-transmitted pathogens.

粘膜是病原体传播的主要部位，但注射疫苗导致粘膜免疫力较差。 颊组织中的粘膜树突状细胞亚群可以诱导免疫和对抗原的耐受。这 拟议的研究重点是通过激活粘膜树突状细胞来诱导粘膜免疫 重组外膜囊泡（rOMV）。在过去的十年里，我们的研究团队设计并 开发了 rOMV 来诱导针对肽抗原的强大 Th1/Th2 免疫反应。在最近的时期 在未发表的工作中，我们的团队发现了一种激活和推动 DC 成熟的方法。也许大多数 令人兴奋的是，我们量化了 rOMV 在猪颊粘膜组织中的渗透深度，并显示 ~100 nm 囊泡穿透组织的深度与幼稚骨髓树突状细胞群一致 细胞（>100 µm）。因此，这种基于 rOMV 的技术具有与粘膜递送一致的特征 疫苗，特别是通过口腔组织。研究的目的是阐明和确立独特性 rOMV 方法作为口腔疫苗递送方法的应用。通过将亚基表位的使用与 为了研究 rOMV 的组织穿透性，我们将评估将这些 rOMV 施用到组织中的假设 颊粘膜会诱导幼稚 DC 的激活，并诱导强大且平衡的 Th1/Th2 免疫 回复。将通过完成以下目标来评估该假设。目标 1：量化 对“弱”抗原的免疫反应以确定 rOMV 的功效和效力。我们的 之前的工作使用绿色荧光蛋白（GFP）作为“弱”抗原的有效模型，证明 皮下注射 rOMV 的效力。我们将使用相同的方法来量化体液 以及对由 rOMV 传递到猪颊粘膜的 GFP 的细胞反应（最接近的动物模型） 人类颊粘膜）。结果将与皮下注射的相同制剂进行比较 （即基准），以便将口腔给药途径的功效与传统给药途径相关联 行政。目标 2：量化对流感血凝素 (HA) 抗原的免疫反应，以 确定 rOMV 赋予的保护性免疫力。在之前的工作中，我们使用了一系列 HA 通过向小鼠皮下注射 rOMV 来量化保护性免疫水平（测量结果） 通过血凝素抑制测定和抗体中和分析。我们将使用相同的方法 量化 rOMV 递送至猪颊粘膜的 HA 的体液和细胞反应。 结果将与皮下注射的相同制剂（即基准）进行比较，以便 将口腔给药途径的功效与传统给药途径相关联。康奈尔大学- Putnam（rOMV 疫苗设计）和 Chang（猪免疫）研究团队结合了两个实验室 专家们共同拥有创建有效的口腔给药疫苗输送平台的技能， 可能适用于多种粘膜传播的病原体。