Dissecting the mechanism of age-specific adjuvant synergy in vitro and in vivo

 描述（由申请人提供）：疫苗是一种重要的公共卫生干预措施，但迄今为止，其开发主要是临时性和经验性的。感染最常见于年龄的极端，疫苗佐剂可能是增强具有不同免疫力的特殊人群免疫原性的关键方法。佐剂，增强免疫反应的分子，可以大大提高免疫力，但仍有很多关于佐剂作用，可以随年龄显着变化的了解。为了鉴定有前景的佐剂系统并表征其作用机制，Levy实验室建立了新型人体外模型系统，包括全血测定、单核细胞衍生的树突细胞（MoDC）阵列和微生理三维组织构建体，其模拟先天性和适应性免疫应答。在使用人类新生儿、婴儿和成人白细胞的初步体外研究中，我们已经鉴定了某些Toll样受体激动剂（TLRA）和C型凝集素受体激动剂（CLRA）组合，其协同增强早期生命中的NF-κ B和炎性小体活化以及Th 1细胞因子产生：合成咪唑并喹啉TLR 7/8激动剂R848和Mincle（NH3）激动剂海藻糖-6，6-二山嵛酸酯（TDB）。在拟定研究中，我们将与Peter Andersen博士（Statens Serum Institut（SSI）;哥本哈根，丹麦）合作，其实验室在佐剂制剂和疫苗学方面拥有世界一流的专业知识，以进一步表征使用呼吸道合胞病毒（RSV）融合前蛋白作为模型抗原的这种组合佐剂系统的协同活性的机制。我们的假设是TLRA和CLRA协同激活NF-κB和炎性体途径，在体外和体内诱导强大的Th 1免疫应答。我们的目标是使用我们的新型人类体外平台来验证这一假设 以及我们的佐剂组合在使用RSV融合前蛋白作为模型抗原中的体内安全性和免疫原性。我们将通过追求以下具体目标（SA）来实现这一目标：目标1：表征组合佐剂协同作用在体外活化人DC中的机制。目的2：评估联合佐剂在体外增强DC介导的淋巴细胞活化的能力。目的3：评价组合佐剂对疫苗反应原性和免疫原性生物标志物的影响。总的来说，这些特定目标的成功追求将为体外和体内年龄特异性佐剂协同作用的机制提供新的见解，从而推进新的方法来增强特殊人群（包括非常年轻的人群）对关键病原体的保护性免疫应答。

 DESCRIPTION (provided by applicant): Vaccines are a crucial public health intervention, yet to date their development has been largely ad hoc and empiric. Infections are most common at the extremes of age and vaccine adjuvantation can be a key approach to enhance immunogenicity for special populations that have distinct immunity. Adjuvants, molecules that boost immune response, can greatly enhance immunity but much remains to be learned regarding adjuvant action that can vary markedly with age. To identify promising adjuvantation systems and characterize their mechanisms of action, the Levy Lab has established novel human in vitro model systems, including whole- blood assays, monocyte-derived dendritic cell (MoDC) arrays, and microphysiologic three-dimensional tissue constructs, which model innate and adaptive immune responses. In preliminary in vitro studies employing human newborn, infant and adult leukocytes, we have identified certain Toll-like receptor agonist (TLRA) and C-type lectin receptor agonist (CLRA) combinations that synergistically enhance NF-kB and inflammasome activation as well as Th1 cytokine production in early life: the synthetic imidazoquinoline TLR7/8 agonist R848 and the Mincle (CLR) agonist Trehalose-6,6-dibehenate (TDB). In the propose studies, we will work in collaboration Dr. Peter Andersen (Statens Serum Institut (SSI); Copenhagen, Denmark), whose laboratory has world class expertise in adjuvant formulation and vaccinology, to further characterize the mechanisms underlying the synergistic activity of this combination adjuvantation system using Respiratory Syncitial Virus (RSV) pre-fusion protein as a model antigen. Our hypothesis is that TLRAs and CLRAs synergistically activate the NF-κB and inflammasome pathways to induce robust Th1 immune responses in vitro and in vivo. Our goal is to test this hypothesis employing our novel human in vitro platforms and in vivo safety immunogenicity of our adjuvant combination in using RSV pre-fusion protein as model antigen. We will achieve this goal by pursuing the following Specific Aims (SAs): Aim 1: Characterize mechanisms of combination adjuvant synergy in activating human DCs in vitro. Aim 2: Assess the ability of combination adjuvants to enhance DC-mediated lymphocyte activation in vitro. Aim 3: Evaluate the impact of combinatorial adjuvants on biomarkers of vaccine reactogenicity and immunogenicity in vitro and in vivo. Overall, successful pursuit of these Specific Aims will provide fresh insight into mechanisms of age-specific adjuvant synergy in vitro and in vivo thereby advancing new approaches to enhance protective immune responses in special populations, including the very young, against key pathogens.