Mechanisms of dmLT Adjuvant

dmLT 佐剂的作用机制

• 批准号: 10066139
• 负责人: Elizabeth B Norton
• 金额: $ 26.07万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-02 至 2021-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10066139
• 关键词: Adjuvant; Adjuvanticity; Antibodies; Antigen Presentation; Antigen-Presenting Cells; Antigens; Automobile Driving; Bacterial Antigens; Binding; Biological Markers; CD86 gene; CREB1 gene; Cell Line; Cell model; Cellular Metabolic Process; Chemicals; Clinical; Clinical Research; Clinical Trials; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Data; Diarrhea; Disease; Dose; Enterotoxins; Epithelial Cells; Escherichia coli Vaccines; Exposure to; Family; Formulation; Genetic Transcription; Genus Hippocampus; Glycolysis; Goals; Health; Human; ITGAX gene; Immune response; Immunity; Immunization; Immunoglobulin A; Immunologics; Inflammasome; Interleukin-1 beta; Interleukin-6; Knock-out; Link; Longevity; Mediating; Medical Research; Memory; Metabolic; Metabolism; Modeling; Molecular; Mucosal Immune Responses; Mucosal Immunity; Mucous Membrane; Mus; Mutation; Oral; Outcome; Oxidative Phosphorylation; PPAR gamma; Parents; Pathway Analysis; Peripheral Blood Mononuclear Cell; Phase; Phase I Clinical Trials; Phenotype; Poliomyelitis; Property; Proteins; Public Health; Research; Research Personnel; Residual state; Role; Safety; Sampling; Serological; Serum; Shigella; Signal Pathway; Signal Transduction; Surface; T cell response; T-Cell Activation; TNFRSF5 gene; Testing; Toxic effect; Up-Regulation; Vaccination; Vaccine Adjuvant; Vaccine Antigen; Vaccine Clinical Trial; Vaccines; Viral Antigens; base; clinically relevant; cost effective; cytokine; design; early detection biomarkers; enterotoxigenic Escherichia coli; fatty acid oxidation; functional outcomes; improved; in vivo; inhibitor/antagonist; metabolic phenotype; metabolic profile; metabolomics; microorganism; monocyte; mouse model; novel; novel vaccines; pre-clinical; predicting response; prevent; response; treatment comparison; vaccine development; vaccine efficacy; vaccine response; vaccine trial

As the majority of infectious microorganisms either colonize or cross mucosal surfaces to enter the host, development of vaccines that induce protective, lasting mucosal immune responses is a crucial public health strategy. dmLT is a promising adjuvant from the heat-labile enterotoxin (LT)-based adjuvant family. It has demonstrated a unique ability to promote mucosal immunity, dose-sparing, and response longevity after parenteral or mucosal immunization in pre-clinical and early phase clinical studies. The interplay between toxicity and adjuvanticity has often complicated a clear perspective on the immunologic and molecular mechanisms occurring with LT-based adjuvants as well as their use clinically. With dmLT’s current safety and efficacy record, it is the best candidate adjuvant from this family. dmLT also provides a unique opportunity to determine the molecular mechanisms of adjuvanticity. As vaccine development moves toward design of more sophisticated, rational, and tailored immunologic responses, understanding the mechanisms of any vaccine component, like dmLT adjuvant, is a primary concern. The objective of this proposal is to define the key molecular mechanisms of dmLT and early biomarkers of adjuvant activity. It has been established that adjuvanticity of LT and dmLT involves activation of cAMP, PKA and the inflammasome in antigen presenting cells (APCs); but no downstream transcriptional effectors, metabolic changes or comprehensive signaling pathway leading to cellular activation have been identified. Our preliminary data indicates that dmLT has multiple signaling responses occurring simultaneously to stimulate antigen-presenting cells to promote vaccine responses. These signaling responses, to which both the A- and B-subunit contribute, are similar to LT in APCs, but not epithelial cells. In the current proposal, we will use our unique arsenal of LT-related proteins, sophisticated omics analyses, and relevant clinical trial samples to definitively answer key mechanistic questions. These include: (1) What are the molecular mechanisms of adjuvanticity of dmLT within antigen presenting cells? (2) Is altered cellular metabolism required for adjuvant stimulation? (3) Using clinical trial samples, does the magnitude of dmLT-induced signaling or serological indicators 24h post-vaccination predict vaccination outcomes? Upon completion of these aims, we expect to generate a much deeper understanding of the major molecular mechanisms of dmLT adjuvant, thereby supporting optimal and rational vaccine development. In addition, we will provide novel information regarding the involvement of host cell metabolism, which has yet to be explored for vaccine adjuvants. This information will further our long-term goal, to identify and utilize the unique properties of dmLT adjuvant within vaccines for the benefit of human health.

由于大多数传染性微生物要么定植或穿过粘膜表面进入宿主， 开发诱导保护性、持久的粘膜免疫反应的疫苗对于公共卫生至关重要 dmLT 是一种基于不耐热肠毒素 (LT) 佐剂家族的有前途的佐剂。 具有促进粘膜免疫、节省剂量和延长反应时间的独特能力 临床前和早期临床研究中的肠胃外或粘膜免疫之间的相互作用。 毒性和佐剂性常常使免疫学和分子生物学的清晰视角变得复杂化。 基于 LT 的佐剂发生的机制及其临床应用。 功效记录，它是该家族的最佳候选佐剂，也提供了独特的机会。 随着疫苗开发朝着更多设计方向发展，确定佐剂的分子机制。 复杂、合理、量身定制的免疫反应，了解任何疫苗的机制 诸如 dmLT 佐剂之类的成分是主要关注点。该提案的目标是定义关键。 dmLT 的分子机制和佐剂活性的早期生物标志物。 已经确定 LT 和 dmLT 的佐剂作用涉及 cAMP、PKA 和 抗原呈递细胞 (APC) 中存在炎症体，但没有下游转录效应子、代谢因子 我们已经确定了导致细胞激活的变化或综合信号通路。 初步数据表明 dmLT 具有同时发生的多种信号反应以刺激 抗原呈递细胞促进疫苗反应，A- 和 A- 均会产生这些信号反应。 B 亚基的贡献与 APC 中的 LT 类似，但与上皮细胞不同。在当前提案中，我们将使用我们的。 独特的 LT 相关蛋白库、复杂的组学分析以及相关的临床试验样本 明确回答关键的机制问题，包括：（1）其分子机制是什么。 (2)佐剂是否需要改变细胞代谢 (3) 使用临床试验样本，dmLT 诱导的信号传导或血清学的强度是否 疫苗接种后 24 小时指标可预测疫苗接种结果吗？ 完成这些目标后，我们期望对主要分子有更深入的了解 此外，我们还研究了 dmLT 佐剂的作用机制，从而支持最佳和合理的疫苗开发。 将提供有关宿主细胞代谢参与的新信息，这一点还有待探索 这些信息将进一步推动我们的长期目标，即识别和利用其独特特性。 在疫苗中添加 dmLT 佐剂有利于人类健康。

项目成果

LTA1 and dmLT enterotoxin-based proteins activate antigen-presenting cells independent of PKA and despite distinct cell entry mechanisms.

尽管细胞进入机制不同，LTA1 和 dmLT 肠毒素蛋白仍能独立于 PKA 激活抗原呈递细胞。

• DOI: 10.1371/journal.pone.0227047
• 发表时间: 2020
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Valli,Eduardo;Baudier,RobinL;Harriett,AmandaJ;Norton,ElizabethB
• 通讯作者: Norton,ElizabethB

Fentanyl conjugate vaccine by injected or mucosal delivery with dmLT or LTA1 adjuvants implicates IgA in protection from drug challenge.

• DOI: 10.1038/s41541-021-00329-0
• 发表时间: 2021-05-13
• 期刊: NPJ vaccines
• 影响因子: 9.2
• 作者: Stone AE;Scheuermann SE;Haile CN;Cuny GD;Velasquez ML;Linhuber JP;Duddupudi AL;Vigliaturo JR;Pravetoni M;Kosten TA;Kosten TR;Norton EB
• 通讯作者: Norton EB

Elevated Extracellular cGMP Produced after Exposure to Enterotoxigenic Escherichia coli Heat-Stable Toxin Induces Epithelial IL-33 Release and Alters Intestinal Immunity.

• DOI: 10.1128/iai.00707-20
• 发表时间: 2021-03-17
• 期刊: Infection and immunity
• 影响因子: 3.1
• 作者: Motyka NI;Stewart SR;Hollifield IE;Kyllo TR;Mansfield JA;Norton EB;Clements JD;Bitoun JP
• 通讯作者: Bitoun JP

Route and antigen shape immunity to dmLT-adjuvanted vaccines to a greater extent than biochemical stress or formulation excipients.

• DOI: 10.1016/j.vaccine.2023.01.033
• 发表时间: 2023-02-24
• 期刊: VACCINE
• 影响因子: 5.5
• 作者: Stone, Addison E.;Rambaran, Saraswatie;V. Trinh, Ivy;Estrada, Marcus;Jarand, Curtis W.;Williams, Blake S.;Murrell, Amelie E.;Huerter, Chelsea M.;Bai, William;Palani, Surya;Nakanishi, Yukihiro;Laird, Renee M.;Poly, Frederic M.;Reed, Wayne F.;White, Jessica A.;Norton, Elizabeth B.
• 通讯作者: Norton, Elizabeth B.