Combination Adjuvants to Activate Human Dendritic Cell Subsets and B Cells

激活人树突状细胞亚群和 B 细胞的组合佐剂

• 批准号: 10162208
• 负责人: Jacques F Banchereau
• 金额: $ 14.17万
• 依托单位: JACKSON LABORATORY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-30 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10162208
• 关键词: 2019-nCoV; ATAC-seq; Accounting; Adjuvant; Antibodies; Antibody Response; Antigen-Presenting Cells; Antigens; B-Lymphocytes; Biological Assay; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; COVID-19; COVID-19 vaccine; CRISPR/Cas technology; Cells; Collaborations; Combined Vaccines; Communicable Diseases; Dendritic Cells; Elderly; Epigenetic Process; Exposure to; FDA approved; Failure; Genes; Genetic Transcription; Glycoproteins; Goals; Helper-Inducer T-Lymphocyte; Human; Humoral Immunities; Immune response; Immune system; Immunity; Immunocompromised Host; Immunoglobulin A; Immunoglobulin G; Immunologic Receptors; In Vitro; Inactivated Vaccines; Individual; Industry; Investigation; Methods; Middle East Respiratory Syndrome Coronavirus; Molecular; Morbidity - disease rate; Mus; Output; Pathway interactions; Plant Roots; Population; Proteomics; Public Health; Recombinants; Research; Research Personnel; SARS coronavirus; Safety; Subunit Vaccines; T cell response; T-Lymphocyte; Technology; The Jackson Laboratory; Therapeutic; Vaccination; Vaccine Adjuvant; Vaccine Antigen; Vaccines; Work; bioinformatics infrastructure; combat; computer infrastructure; design; epigenomics; humanized mouse; immunogenicity; immunosuppressed; in vivo; innovation; knock-down; metabolomics; mortality; mouse model; pandemic disease; pathogen; prevent; programs; public health relevance; reconstitution; response; side effect; tool; transcriptomics

PROJECT SUMMARY Vaccination is the most effective method for preventing infectious diseases. Many current vaccines are “inactivated” or “subunit” vaccines composed of purified or recombinant pathogen components to which an adjuvant is often added to increase the magnitude of antibody responses. However, subunit vaccines formulated with current FDA-approved adjuvants do not sufficiently boost immunity in some populations, particularly immunocompromised and elderly subjects. Numerous adjuvants have been discovered in recent years and show enhanced immunogenicity as single agents; however, little is known about the activity of their combination, their safety, their efficacy and their mechanisms of action. Responses to vaccination and adjuvants involve dendritic cells (DCs), which capture and present vaccine antigens thereby facilitating the differentiation of follicular helper T cells (Tfh) and B cells and subsequent humoral immunity. Therefore, we propose to examine the molecular mechanisms and functional outputs of human DC subsets exposed to combination adjuvants ex vivo and in vivo. The focus on human DCs is essential given the substantial differences in innate immune receptor distribution and function between the mouse and the human. Our goal is to select a combination adjuvant using functional assays, followed by in-depth investigation of molecular pathways accounting for enhanced immunogenicity. Our collaboration with industry will enable the transition of the selected combination adjuvant to further studies of human vaccination. Our Specific Aims are built towards this goal. Thus, first we will screen adjuvant combinations by assessing the capacity of adjuvant-activated human DC subsets to skew the differentiation of naïve CD4+T cells into Tfh cells that secrete IL-21 and induce B cells to produce IgG and IgA antibodies (Aim 1). Promising combinations will be further studied in DCs using validated, sensitive and high-throughput transcriptomic epigenomic, proteomic and metabolomic methods, and by functional knockdown in vitro, to determine the underlying molecular pathways of adjuvant efficacy (Aim 2). We will then validate the identified molecular pathways through functional knockdown studies in vivo using humanized mice carrying a functionally reconstituted human immune system, which will also enable the examination of possible side effects (Aim 3). The proposed research program will leverage cutting-edge epigenetic (ATAC-seq), transcriptional, gene editing (CRISPR/Cas9) and metabolomic technologies; innovative humanized mouse models; a powerful computational and bioinformatics infrastructure at The Jackson Laboratory; and the complementary expertise of a dynamic team of investigators. Our deliverable is a combination adjuvant for enhanced humoral immunity and molecular pathways that are essential for its efficacy.

项目概要 疫苗接种是预防传染病最有效的方法。 由纯化或重组病原体成分​​组成的“灭活”或“亚单位”疫苗 通常添加佐剂以增加抗体反应的强度，但是，配制的亚单位疫苗。 目前 FDA 批准的佐剂不足以增强某些人群的免疫力，特别是 近年来发现了许多佐剂并表明其适用于免疫功能低下和老年受试者。 作为单一药物增强免疫原性；然而，对其组合的活性、其活性知之甚少。 安全性、有效性及其作用机制 对疫苗和佐剂的反应涉及树突状。 细胞（DC），捕获并呈递疫苗抗原，从而促进滤泡辅助细胞的分化 T细胞（Tfh）和B细胞以及随后的体液免疫因此，我们建议检查分子。 暴露于离体和体内组合佐剂的人类 DC 亚群的机制和功能输出。 鉴于先天免疫受体分布的巨大差异，对人类 DC 的关注至关重要 我们的目标是使用功能来选择组合佐剂。 分析，然后深入研究增强免疫原性的分子途径。 与工业界的合作将使选定的组合佐剂过渡到进一步研究 我们的具体目标就是为了实现这一目标，因此，首先我们将筛选佐剂组合。 通过评估佐剂激活的人类 DC 亚群扭曲初始 CD4+T 分化的能力 细胞转化为 Tfh 细胞，分泌 IL-21 并诱导 B 细胞产生 IgG 和 IgA 抗体（目标 1）。 将使用经过验证的、灵敏的和高通量的转录组学在 DC 中进一步研究组合 表观基因组学、蛋白质组学和代谢组学方法，并通过体外功能敲低，确定 然后我们将验证佐剂功效的潜在分子途径（目标 2）。 使用携带功能性基因的人源化小鼠进行体内功能敲低研究 重建人体免疫系统，这也将能够检查可能的副作用（目标 3）。 拟议的研究计划将利用尖端的表观遗传学（ATAC-seq）、转录、基因编辑 (CRISPR/Cas9) 和代谢组学技术；创新的人源化小鼠模型； 杰克逊实验室的生物信息学基础设施以及动态的互补专业知识； 我们的研究成果是增强体液免疫和分子免疫的组合佐剂。 对其功效至关重要的途径。