Determining the Molecular Mechanism of Covalently Linked TLR Agonists

 描述（由申请方提供）：许多病毒疫苗通过模式识别受体的协同组合刺激免疫系统。这些激动剂的组合对于长寿命的抗体和T细胞应答是关键的。这些多激动剂反应可用作新的佐剂，但它们的作用途径和机制仍不清楚。我们的目标是阐明佐剂协同作用的机制途径，该途径是由抗原呈递细胞上的两种共价连接的Toll样受体（TLR）激动剂（Dual_TLR）对两种受体的分子活化引起的。我们将确定一系列协同作用的位置及其发生的机制。我们将使用模型疫苗测试我们的协同佐剂的安全性和有效性。我们将使用合成化学和分子生物学工具的组合来探测两个TLR协同激活的物理和生物学机制。在本申请中，我们研究了连接在一起的两种TLR激动剂的增强活性和抗体应答。先前，我们表明连接不同TLR（例如TLR2_TLR9）的激动剂导致增加的激活。用Dual_TLR激动剂作为佐剂的疫苗改善了抗体应答的广度和强度。此外，这些协同作用由接头长度和受体身份控制，这意味着通过物理现象在分子水平上进行控制。研究Dual_TLR激动剂的机制将导致对细胞内信号协同作用的基本理解，以及对将细胞应答与TLR信号介导的抗体多样性联系起来的机制的理解。有了这些基础知识，将更容易将从病毒疫苗中获得的知识应用于设计新的佐剂。

 DESCRIPTION (provided by applicant): Many viral vaccines stimulate the immune system through a synergistic combination of pattern recognition receptors. These combinations of agonists are critical for long-lived antibody and T- cell responses. These multi-agonist responses could be applied as new adjuvants, but the pathways and mechanisms through which they operate remain unclear. Our goal is to elucidate a mechanistic pathway for adjuvant synergies resulting from the molecular activation of two receptors by two covalently linked Toll-like Receptor (TLR) agonists (Dual_TLRs) on antigen presenting cells. We will determine the location of a set of synergies and the mechanism by which they occur. We will test the safety and efficacy of our synergistic adjuvants using model vaccines. We will use a combination of synthetic chemistry and molecular biology tools to probe physical and biological mechanisms for the synergistic activation of two TLRs. In this application, we study the enhanced activity and antibody response of two TLR agonists that are linked together. Previously, we showed that linking agonists of distinct TLRs (e.g. TLR2_TLR9) leads to increased activation. Vaccines adjuvanted with Dual_TLR agonists improve the breadth and intensity of antibody response. Moreover, these synergies are controlled by the linker-length and receptor identity implying control at the molecular level by physical phenomenon. Studying the mechanism of Dual_TLR agonists will result in fundamental understanding of signaling synergies within cells as well as an understanding of the mechanism that links cellular responses to antibody diversity mediated by TLR signaling. With this fundamental knowledge, it will be easier to apply the knowledge gained from viral vaccines to the designing new adjuvants.