Mechanistic evaluation of mast cell agonists combined with TLR, NOD and STING agonists.

肥大细胞激动剂联合 TLR、NOD 和 STING 激动剂的机制评估。

• 批准号: 10657847
• 负责人: Kristy M Ainslie
• 金额: $ 64.52万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-03 至 2027-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10657847
• 关键词: Acids; Adjuvant; Agonist; Anaphylaxis; Animal Model; Antigen-Presenting Cells; Antigens; Area; Attenuated; Autoimmunity; Autopsy; Binding; Biocompatible Materials; Blood Chemical Analysis; CD8-Positive T-Lymphocytes; Categories; Cell Degranulation; Cells; Cold Chains; Contracts; Data; Delayed Hypersensitivity; Dextrans; Dose; Elderly; Emulsions; Encapsulated; Ensure; Environment; Evaluation; Formulation; Genes; Genetic; Genetic Variation; Goals; Histamine; Histology; Human; Hydrophobicity; IgE; Immune; Immune response; Immune system; Immunity; Immunization; Immunohistochemistry; Immunologic Stimulation; In Vitro; Inbred Strains Mice; Infection; Knockout Mice; Langerhans cell; Leukocytes; Leukotrienes; Lipid A; Liposomes; Lymphocyte; Maps; Measures; Memory; Memory B-Lymphocyte; Methods; Modeling; Mouse Strains; Mus; Nucleotides; Ovalbumin; Pathology; Pattern recognition receptor; Peptides; Population; Population Heterogeneity; Predisposition; Production; Prostaglandins; Proteins; Receptor Cell; Recombinants; Reporting; Resources; Safety; Schedule; Serum; Severity of illness; Site; Smallpox; Stimulator of Interferon Genes; Subunit Vaccines; T cell response; TLR2 gene; Toll-like receptors; Toxic effect; Tryptase; United States National Institutes of Health; Vaccination; Vaccine Research; Vaccines; Vaccinia; Vaccinia Vaccine; Viral; Work; adaptive immune response; aged; aging population; aluminum sulfate; antigen test; antigen-specific T cells; controlled release; cytokine; cytotoxicity; draining lymph node; human model; in vivo; insight; mast cell; monocyte; mouse model; particle; pathogen; protective effect; recruit; response; small molecule; synergism; tool; vaccine efficacy; vaccine evaluation; vaccine formulation

ABSTRACT Subunit vaccines are safer and can more broadly be applied across the population then other vaccine formulations such as live-attenuated. However, subunit antigens are often poorly antigenic and require formulation with an immune stimulating adjuvant to garner protection. Additionally, some vaccines require more than one adjuvant, necessitating combined adjuvants to stimulate a protective response. Also, a combined adjuvant could decrease vaccination boosts and provide longer protection. One avenue to evaluate combined adjuvants is with mast cell (MC) agonists. MCs are throughout the body and reside at many interfaces of the host and the environment. When activated MCs recruit monocytes and leukocytes to the local area and help to promote an adaptive response. MC agonists combined with toll-like receptor (TLR), nucleotide-binding oligomerization domain-containing protein 2 (NOD-2), or stimulators of interferon genes (STING) agonists should elicit not only a humoral response, but also a cellular response to create an efficacious and effective vaccine. Herein we will evaluate combined MC agonists with TLR, NOD-2 or STING agonist to identify synergistic pairs. Pairs will be evaluated in mice and human cells as well as with cells from collaborative cross (CC) strains. The CC strains are a large panel of recombinant inbred mouse strains with genetic variation that can mimic the human population as well as give insight into genetic variables that contribute to adjuvant mechanism. To ensure that the adjuvants are co-delivered as well as offer dose sparring, storage outside the cold chain and controlled release of adjuvant, we will formulate them into acetalated dextran (Ac-DEX) microparticles. Ac-DEX formulations have illustrated enhanced delivery of STING, NOD-2, TLR and MC agonists in vitro and in vivo, above that of other carriers like liposomes or PLGA particles. The best identified adjuvant combination will be evaluated in a mouse model of a vaccinia vaccine with subunit antigen BR8. We will use pattern recognition receptor (PRR) knock-out mice as well as cell deficient mice to elucidate aspects of the combination adjuvant's mechanism. Additionally, we will employ genetic sequencing tools to mechanistically identify the combination adjuvants mechanism.

摘要 与其他疫苗相比，亚单位疫苗更安全，可以更广泛地应用于人群 活体减毒等制剂。然而，亚单位抗原通常抗原性较差，需要 含有免疫刺激佐剂的配方以获得保护。此外，一些疫苗需要更多 而不是一种佐剂，需要联合佐剂来刺激保护性反应。此外，合并后的 佐剂可以减少疫苗接种次数，并提供更长时间的保护。综合评价的一种途径 佐剂使用肥大细胞(MC)激动剂。MCS遍布全身，驻留在 主机和环境。当被激活时，MC将单核细胞和白细胞招募到局部区域，并帮助 促进适应性反应。MC激动剂与Toll样受体(TLR)结合，核苷酸结合 低聚含结构域蛋白2(NOD-2)或干扰素基因刺激物(STIN)激动剂应 不仅引起体液反应，还引起细胞反应，以创造出有效和有效的疫苗。 在这里，我们将评估MC激动剂与TLR、NOD-2或STING激动剂的联合作用，以确定协同作用对。 配对将在小鼠和人类细胞以及来自协作杂交(CC)菌株的细胞中进行评估。这个 CC品系是一大批具有遗传变异的重组近交系小鼠品系，可以模仿 并提供了对佐剂机制有贡献的遗传变量的见解。为了确保 佐剂是联合递送的，并提供剂量辩论、冷链之外的储存和受控 释放佐剂后，我们将它们制成乙缩醛葡聚糖(Ac-DEX)微粒。AC-DEX 制剂表明，在体外和体内，STIN、NOD-2、TLR和MC激动剂的释放增加， 高于其他载体，如脂质体或PLGA颗粒。最好的佐剂组合将是 在含有BR8亚单位抗原的牛痘疫苗的小鼠模型中进行评估。我们将使用模式识别 受体(PRR)敲除小鼠以及细胞缺陷小鼠，以阐明联合佐剂的某些方面 机制。此外，我们将使用基因测序工具来机械地识别这种组合 佐剂机制。