Mechanisms of Nanomaterials-based Combination Adjuvants

项目摘要 模式识别受体（PRRs）的发现，包括Toll样受体（TLR）和NOD样受体（NOD样受体）， 受体（NLR）的研究导致了对佐剂制剂中先天免疫的分子激动剂的研究。 一个新兴的范例是仔细选择佐剂组合可以导致互补的甚至是互补的。 协同增强疫苗诱导的免疫应答。大多数联合佐剂 研究是与PRR激动剂混合的贮库佐剂的化学非均相混合物， 从批次间的差异性和差的化学定义，使机制和安全性的调查， 挑战.我们的实验室研究自组装肽纳米纤维（PNF）作为疫苗佐剂。一个关键 PNF相对于乳液佐剂的优点是自缔合肽的一级序列可以 设计用于控制PNF的物理化学特征，例如形态、电荷、手性或 疏水性是佐剂活性的关键因素。对作用模式的机械论见解 表明与PAMPs不同，PNF不会引起DC成熟，但促进与PAMPs相关的DAMPs的释放。 渗透/氧化应激。在该应用中，我们提出开发由以下组成的组合佐剂： 化学上确定的DAMP诱导肽纳米纤维（PNF）和TLR 2/NOD 2激动剂。 我们的目标是了解DAMP诱导PNF和PRR激动剂的分子机制 协调先天免疫信号传导并诱导互补、协同或抑制的反应， 平衡免疫原性和安全性。在目的1中，我们将研究具有不同理化性质的PNF的效果。 本发明还涉及TLR 2或NOD 2激动剂组合对DC活化、DAMP释放和抗原呈递的影响。 在目标2中，使用实验设计（DOE）方法，我们将开发一种最佳配方， 控制PNF-TLR 2和PNF-NOD 2的组合，并确定先天性 使用各种KO小鼠模型在DC中进行免疫。在目标3中，我们将验证PNF-TLR 2-NOD 2的功效。 组合佐剂并使用人DC研究翻译潜力。 拟议研究的结果将促进我们对介导的分子机制的理解。 对PNF-PRR激动剂佐剂组合的先天免疫应答，并将导致新的组合- 具有高翻译潜力的用于对抗感染性和非感染性疾病的佐剂平台。

PROJECT SUMMARY The discovery of pattern recognition receptors (PRRs), including toll-like receptors (TLRs) and NOD-like receptors (NLRs) has led to the investigation of molecular agonists of innate immunity in adjuvant formulations. An emerging paradigm is that careful selection of adjuvant combinations can result in complementary and even synergistic enhancement of vaccine-induced immune responses. Most combination adjuvants under investigation are chemically heterogeneous mixtures of depot adjuvants mixed with PRR agonists and suffer from batch-to-batch variability and poor chemical definition making investigation of mechanisms and safety a challenge. Our lab investigates self-assembling peptide nanofibers (PNFs) as vaccine adjuvants. A key advantage of PNFs over emulsion adjuvants is that the primary sequence of the self-associating peptide can be designed to control the physicochemical features of PNFs such as morphology, charge, chirality, or hydrophobicity, which are key contributors to adjuvant activity. Mechanistic insights into the mode of action indicates that unlike PAMPs, PNFs do not cause DC maturation but facilitate the release of DAMPs related to osmotic/oxidative stress. In this application, we propose to develop combination adjuvants composed of chemically defined DAMP-inducing peptide nanofibers (PNFs) and TLR2/NOD2 agonists. Our objectives are to understand how molecular mechanisms of DAMP-inducing PNFs and PRR agonists orchestrate innate immune signaling and induce responses that are complimentary, synergistic, or inhibitory for balancing immunogenicity with safety. In aim 1, we will examine the effect of PNFs with varying physicochemical properties and TLR2 or NOD2 agonist combinations on DC activation, DAMP release, and antigen presentation. In aim 2, using a design of experiments (DOE) approach, we will develop an optimal formulation with precisely controlled PNF-TLR2 and PNF-NOD2 combinations and determine the molecular mechanisms of innate immunity in DCs using various KO mouse models. In aim 3, we will validate the efficacy of PNF-TLR2-NOD2 combination adjuvants and investigate translational potential using human DCs. Outcomes of the proposed studies will advance our understanding of the molecular mechanisms that mediate innate immune responses to PNF-PRR agonist adjuvant combinations and will lead to new combinatorial- adjuvant platforms for combating infectious and non-infectious diseases with high translational potential.