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: 9890987
• 负责人: Jacques F Banchereau
• 金额: $ 60.5万
• 依托单位: JACKSON LABORATORY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-15 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9890987
• 关键词: ATAC-seq; Accounting; Adjuvant; Agonist; Antibodies; Antibody Response; Area; B-Lymphocytes; Biological Assay; CD34 gene; CD4 Positive T Lymphocytes; CRISPR/Cas technology; Cells; Collaborations; Communicable Diseases; Computational Biology; Cytometry; Dendritic Cell Vaccine; Dendritic Cells; Elderly; Elements; Engineering; Engraftment; Epigenetic Process; Event; Exposure to; FDA approved; Flow Cytometry; Genes; Genetic Enhancer Element; Genetic Transcription; Genomics; Goals; Helper-Inducer T-Lymphocyte; Human; Human Engineering; Humoral Immunities; IgA1; IgA2; IgE; IgG1; IgG3; Immune; Immune response; Immune system; Immunity; Immunobiology; Immunocompromised Host; Immunodeficient Mouse; Immunoglobulin A; Immunoglobulin Class Switching; Immunoglobulin G; Immunologic Receptors; In Vitro; Inactivated Vaccines; Individual; Industry; Influenza; Interferon Type II; Interleukin-13; Interleukin-17; Interleukin-4; Investigation; Lead; Metabolic; Methods; Molecular; Morbidity - disease rate; Mus; Outcome; Output; Pathway interactions; Pattern; Phosphorylation; Plasma Cells; Population; Proteomics; Public Health; RNA; Recombinants; Research; Research Personnel; STING agonists; Safety; Selection Criteria; Sendai virus; Signal Transduction; Site; Subunit Vaccines; System; TLR4 gene; TLR7 gene; Technology; Testing; The Jackson Laboratory; Vaccination; Vaccine Antigen; Vaccines; base; bioinformatics infrastructure; computer infrastructure; design; epigenome; epigenomics; genome editing; genomic data; humanized mouse; immunogenicity; immunosuppressed; in vivo; innovation; insight; knock-down; metabolomics; mortality; mouse development; mouse model; novel vaccines; pathogen; prevent; programs; promoter; protein expression; public health relevance; receptor; reconstitution; response; side effect; small hairpin RNA; synergism; tool; transcriptome; transcriptomics; vaccine development; vaccine efficacy

 DESCRIPTION (provided by applicant): 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.

 描述（应用程序提供）：疫苗接种是预防传染病的最有效方法。许多当前的疫苗被“灭活”或“亚基”疫苗组成，该疫苗由纯化或重组的病原体成分​​组成，通常会添加可调节以增加抗体反应的幅度。但是，用当前FDA批准的调节器配制的亚基疫苗在某些人群中没有充分提高免疫力，尤其是免疫功能低下和较旧的受试者。近年来已经发现了许多调节器，并显示出作为单一药物的免疫原性。但是，对它们的组合，安全性，效率和机制的活动知之甚少 行动。对疫苗和调节剂的反应涉及树突状细胞（DC），该细胞捕获并呈现疫苗抗原，从而支持卵泡辅助T细胞（TFH）和B细胞的分化以及随后的人类免疫学。因此，我们建议检查暴露于组合佐剂的人体和体内的人类DC子集的分子机制和功能输出。鉴于小鼠和人之间的先天免疫受体分布和功能存在实质性差异，因此对人类DC的关注至关重要。我们的目标是使用功能评估选择一个组合调整，然后对分子途径进行深入研究，以提高免疫原性。我们与行业的合作将使所选组合的过渡适应人类疫苗接种的进一步研究。我们的具体目标是针对这个目标的。首先，我们将通过评估调整激活的人类DC亚群的能力来筛选调整组合的组合，以使幼稚的CD4+T细胞分化为TFH细胞的分化为TFH细胞，该细胞将IL-21和诱导B细胞产生IgG和IgA抗体（AIM 1）。使用经过验证，敏感和高通量转录组学表观基因组，蛋白质组学和代谢组方法以及通过在体外功能性敲低的情况下，将进一步研究DC，以确定可调节效率的基本分子途径（AIM 2）。然后，我们将使用携带功能重构人类免疫系统的人源化小鼠在体内通过功能敲低研究来验证已识别的分子途径，这也将使可能的副作用检查（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；杰克逊实验室的强大计算和生物信息学基础设施；以及一个充满活力的调查人员团队的补充专业知识。我们的可交付是针对增强的体液免疫学和分子途径的组合，这对于其有效性至关重要。