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.

抽象的 亚基疫苗更安全，可以在整个种群中更广泛地应用于其他疫苗 诸如实时衰减之类的配方。但是，亚基抗原通常抗原较差，需要 具有免疫刺激佐剂的配方，以获得保护。此外，有些疫苗需要更多 比一个佐剂，需要合并的佐剂来刺激保护性反应。另外，合并 佐剂可以减少疫苗接种并提供更长的保护。一条评估合并的途径 佐剂是肥大细胞（MC）激动剂。 MC遍布整个身体，并驻留在 主机和环境。当激活MCS招募单核细胞和白细胞时， 促进自适应反应。 MC激动剂结合了Toll样受体（TLR），核苷酸结合 含寡聚结构域的蛋白2（NOD-2）或干扰素基因（sting）激动剂的刺激剂应应 不仅引起体液反应，还引起细胞反应，以产生有效有效的疫苗。 在此，我们将与TLR，NOD-2或STING激动剂一起评估MC激动剂，以识别协同对。 对小鼠和人类细胞以及协作杂交（CC）菌株的细胞将进行对。这 CC菌株是一大批重组近交小鼠菌株，具有遗传变异，可以模仿 人口以及深入了解有助于辅助机制的遗传变量。确保 佐剂是共同交付的，并提供剂量串联，在冷链外部存放并控制 佐剂的释放，我们将将它们配合到乙酰化的葡萄糖（AC-DEX）微粒中。 AC-DEX 配方表明，在体外和体内增强了Sting，Nod-2，TLR和MC激动剂的递送， 在其他载体（如脂质体或PLGA颗粒）上方的上方。最好的辅助组合将是 用亚基抗原BR8的疫苗疫苗的小鼠模型进行评估。我们将使用模式识别 受体（PRR）敲除小鼠以及细胞不足的小鼠，以阐明组合辅助的方面 机制。此外，我们将采用遗传测序工具来机械地识别组合 佐剂机制。