Molecular Mechanisms of Adjuvant Triplet Combinations

文摘：佐剂三联体组合的分子机制 免疫系统根据微生物输入的复杂组合做出决定。活疫苗 从经验上讲，从病原体中减毒的病毒是产生终身免疫力的有效手段 许多致命的病原体，因为它们模仿对微生物信号组合的免疫反应。然而， 使用佐剂等免疫调节剂的非活疫苗的合理设计仍然是一种 在许多情况下，在活疫苗接种不有效或不可行的情况下，难以完成任务，部分原因是识别有效 佐剂组合和相关的分子机制解释串扰仍然是一个主要的挑战。 根据我们最近的发现和大量的初步数据，我们建议定义分子机制 两种含有Toll样受体(TLR)、C型选择受体的三联体佐剂激动剂 (CLR)、RIG-I样受体(RLR)和胞浆dsDNA传感器(CDS)途径-诱导保护性CD4+和 小鼠的CD8+T细胞反应。我们将使用一种创新的方法，这种方法(I)是比较的-通过对比 佐剂三联体及其配对作为精确定位分子手段的定量效应 解释串扰的机制；以及(Ii)多尺度--通过研究细胞、组织、 还有整个身体。首先，我们将通过以下方法确定细胞内信号串扰的分子机制 通过在佐剂受体近端的蛋白质复合体水平上测试假说， 磷酸化的级联和激酶-底物关系，以及基因调控网络。第二，我们将 确定佐剂组合影响细胞间信号转导的分子机制 树突状细胞(DC)和T细胞通过测试关于形成细胞的调节机制的假说， 树突状细胞表面和分泌的蛋白质组。第三，我们将检验关于佐剂三胞胎对 效应器和记忆性T细胞以及皮肤底层细胞回路在整个生物体内的传播和播种 (疫苗接种部位)和引流淋巴结，这解释了诱导保护性的、长期的T细胞免疫。 这项工作的结果将在佐剂组合研究的前沿产生关键的见解 具有高阶佐剂组合的特征，可以模仿成熟、有效的活体佐剂的效果 减毒疫苗，并为未来针对感染的疫苗设计提供信息。

Abstract: Molecular Mechanisms of Adjuvant Triplet Combinations The immune system makes decisions in response to complex combinations of microbial inputs. Live vaccines that are empirically attenuated from pathogens have been a powerful means to yield life-long immunity against many deadly pathogens because they mimic immune responses to combinations of microbial signals. However, the rational design of non-live vaccines using immunomodulatory agents such as adjuvants has remained an elusive task in many cases where live vaccination is not efficacious or feasible, in part because identifying potent adjuvant combinations and associated molecular mechanisms that explain cross talk remains a major challenge. Based on our recent findings and extensive preliminary data, we propose to define the molecular mechanisms through which two adjuvant triplets – containing agonists for Toll-like receptor (TLR), C-type lection receptor (CLR), RIG-I-like receptor (RLR), and cytosolic dsDNA sensor (CDS) pathways – induce protective CD4+ and CD8+ T cell responses in mice. We will use an innovative approach which is (i) comparative – by contrasting the quantitative effects of adjuvant triplets and matching singles and pairs as means to accurately pinpoint molecular mechanisms explaining cross talk; and (ii) multiscale – by studying molecular mechanisms at play in cells, tissues, and the whole body. First, we will determine the molecular mechanisms of intra-cellular signaling cross talk by adjuvant triplets by testing hypotheses at the level of protein complexes proximal to adjuvant receptors, phosphorylation cascades and kinase-substrate relationships, and gene regulatory networks. Second, we will identify the molecular mechanisms through which adjuvant combinations impact inter-cellular signaling between dendritic cells (DCs) and T cells by testing hypotheses on the regulatory mechanisms shaping the cellular, surface, and secreted proteome of DCs. Third, we will test hypotheses on the effects of adjuvant triplets on the organism-wide spreading and seeding of effector and memory T cells, and the underlying cell circuits of the skin (vaccination site) and draining lymph node that explain the induction of protective, long-term T cell immunity. Results from this work will produce critical insights at the forefront of adjuvant combination research by characterizing higher-order combinations of adjuvants that can mimic the effects of well-established, potent live attenuated vaccines and inform future vaccine designs against infection.