Mechanisms of Nanomaterials-based Combination Adjuvants

项目总结 模式识别受体的发现，包括Toll样受体和NOD样受体 受体(NLR)导致了佐剂制剂中天然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产生互补甚至 协同增强疫苗诱导的免疫反应。以下大多数组合佐剂 调查的是仓库佐剂与PRR激动剂的化学异质混合物，并遭受 从批次到批次的可变性和糟糕的化学定义，使得对机理和安全性的研究成为 挑战。我们的实验室研究了自组装多肽纳米纤维(PNF)作为疫苗佐剂。一把钥匙 与乳剂佐剂相比，PNFS的优点是自结合肽的一级序列可以是 用于控制对氟化钠的物理化学特征，如形态、电荷、手性或 疏水性，它们是佐剂活性的关键贡献者。对行动模式的机械论洞察 表明与PAMP不同，PNP不会导致DC成熟，但会促进与以下相关的湿气的释放 渗透/氧化应激。在本申请中，我们建议开发由以下组成的组合佐剂 化学定义的潮湿诱导多肽纳米纤维(PNF)和TLR2/NOD2激动剂。 我们的目标是了解潮湿诱导的PNFS和PRR激动剂的分子机制 协调先天免疫信号，并诱导对以下方面具有补充、协同或抑制作用的反应 平衡免疫原性和安全性。在目标1中，我们将检查具有不同物理化学特性的PNFS的效果 特性和TLR2或NOD2激动剂组合对DC激活、潮湿释放和抗原提呈的影响。 在目标2中，使用实验设计(DOE)方法，我们将开发出一种具有精确 控制PNF-TLR2和PNF-NOD2结合并确定先天致病的分子机制 不同KO小鼠模型树突状细胞的免疫功能。在目标3中，我们将验证PNF-TLR2-NOD2的有效性 联合佐剂，并利用人类DC研究翻译潜力。 拟议的研究结果将促进我们对调解的分子机制的理解 PNF-PRR激动剂佐剂组合的先天免疫反应，并将导致新的组合- 具有很高翻译潜力的抗击传染病和非传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.