Mechanisms of dmLT Adjuvant

由于大多数有感染性的微生物要么定居，要么穿过粘膜表面进入宿主， 开发能够诱导保护性、持久的粘膜免疫反应的疫苗是一个至关重要的公共卫生问题 策略。DmLT是基于不耐热肠毒素(LT)的佐剂家族中一种很有前途的佐剂。它有 显示出独特的能力，以促进黏膜免疫，节省剂量，并延长反应寿命后 临床前和早期临床研究中的肠外或粘膜免疫。两者之间的相互作用 毒性和佐剂往往使免疫学和分子生物学的清晰前景复杂化。 以LT为基础的佐剂的发生机制及其临床应用。以dmlt目前的安全性和 疗效记录，是该家族最好的候选佐剂。DmLT还提供了一个独特的机会 确定佐剂的分子机制。随着疫苗开发朝着设计更多 复杂、合理和量身定制的免疫反应，了解任何疫苗的机制 成分，如dmLT佐剂，是一个主要问题。这项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。 已证实LT和dmLT的佐剂性涉及cAMP、PKA和 抗原提呈细胞(APC)中的炎症体；但没有下游的转录效应因子，代谢 导致细胞激活的变化或全面的信号通路已经被确定。我们的 初步数据表明，dmLT有多个信号反应同时发生以刺激 抗原提呈细胞促进疫苗反应。这些信号反应，A-和 B亚基的贡献，在APC中类似于LT，但不是上皮细胞。在当前的提案中，我们将使用我们的 独特的LT相关蛋白质、复杂的组学分析和相关的临床试验样本 明确回答关键的机械问题。这些问题包括：(1)分子机制是什么？ 抗原提呈细胞内dmLT的佐剂性？(2)佐剂所需的细胞代谢改变 刺激？(3)使用临床试验样本，dmLT诱导的信号或血清学信号的大小 疫苗接种后24小时指标预测疫苗接种结果？ 在完成这些目标后，我们希望对主要分子有更深入的了解 DmLT佐剂的作用机制，从而支持最佳和合理的疫苗开发。此外，我们 将提供有关宿主细胞新陈代谢的新信息，这还有待探索。 用于疫苗佐剂。这些信息将促进我们的长期目标，识别和利用独特的性质 在疫苗中添加dmLT佐剂，以造福人类健康。

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.