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.

项目摘要 疫苗接种是预防传染病的最有效方法。当前许多疫苗是 由纯化或重组的病原体成分​​组成的“灭活”或“亚基”疫苗 通常添加添加剂以增加抗体反应的大小。但是，配制了亚基疫苗 使用当前FDA批准的调节器并不能充分提高某些人群的免疫力，尤其是 免疫功能低下的主体。近年来发现了许多调节器 作为单个药物的免疫原性增强；但是，对它们的组合的活动知之甚少 安全，效率和行动机制。对疫苗接种和调节器的反应涉及树突状 细胞（DC），捕获和呈现疫苗抗原，从而支持卵泡助手的分化 T细胞（TFH）和B细胞以及随后的体液免疫力。因此，我们建议检查分子 暴露于组合调节器的人体和体内的人类直流子集的机制和功能输出。 鉴于先天免疫接收器分布存在实质性差异，对人类DC的关注至关重要 小鼠和人之间的功能。我们的目标是使用功能选择组合调整 测定，随后对分子途径进行了深入研究，这些途径考虑了增强的免疫原性。我们的 与行业的合作将使选定组合的过渡适应进一步研究 人类疫苗接种。我们的具体目标是针对这个目标的。首先，我们将筛选调整组合 通过评估调整激活的人类DC子集的能力偏差幼稚的CD4+T的分化 细胞进入分泌IL-21并诱导B细胞产生IgG和IgA抗体的TFH细胞（AIM 1）。有希望的 使用经过验证，敏感和高通量转录组的DC将进一步研究DC 表观基因组，蛋白质组学和代谢组方法，并通过在体外功能敲低以确定 可调效率的基本分子途径（AIM 2）。然后，我们将验证已鉴定的分子 使用携带功能的人源化小鼠在体内通过功能敲低研究的途径 重建的人类免疫学系统，这也将能够检查可能的副作用（AIM 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）和代谢组技术；创新的人性化小鼠模型；强大的计算 和杰克逊实验室的生物信息学基础设施；以及动态的完整专业知识 调查人员团队。我们可交付的是调整的组合，以增强体液免疫力和分子 对其效率至关重要的途径。