Directing the Immune System via Polymeric Combinations of Molecular Signals

描述(申请人提供)：该项目旨在通过寻找分子激动剂的协同组合来开发刺激树突状细胞的新方法。长期目标是使用一种指导免疫系统的化学策略来改善疫苗的性能。今天，疫苗仍然是最有效的疾病预防形式。许多疾病的疫苗仍然非常接近现实，但缺乏足够高的有效性来部署。直到最近，合理地改进疫苗还被认为是不可能的，因为疫苗的研制在很大程度上是一个经验过程。现在已经确定了疫苗激活的细胞途径--树突状细胞。这些细胞使用一系列受体产生对疫苗的免疫力，这些受体是由外来病原体携带的分子激动剂刺激的。最有效的疫苗都以协同组合的方式刺激这些受体。一些疫苗使用选定的分子激动剂组合，引导树突状细胞诱导细胞毒素免疫反应或持续的体液反应。这些分子激动剂在最有效的疫苗上的放置和浓度是产生强大免疫反应的关键因素。许多较弱的疫苗没有利用这些协同效应。确定激动剂之间协同作用的方法可能会改进弱疫苗，并揭示关于免疫系统激活的基本知识。然而，目前还没有化学方法来操纵这些分子信号，了解它们的协同效应，或者将它们附加到潜在的疫苗上以增强免疫力。我们建议开发一种通过一系列生物偶联反应在惰性聚合物支架上结合分子激动剂的方法。我们将使用这些激动剂的组合来研究和了解它们在刺激树突状细胞方面的协同活性。我们的假设是，在我们的支架的推动下，不同的激动剂组合将引导免疫系统产生更强的体液免疫或细胞毒免疫。我们还将开发将我们的免疫导向聚合物连接到开发的疫苗上的方法，例如产肠毒素大肠杆菌，这些疫苗对于商业部署来说还不够有效。我们将使用异遥螺旋聚合物合成和一系列生物共轭反应来创建一个激动剂的组合库，这些激动剂以树突状细胞上负责免疫刺激的许多Toll样和Nod样受体为靶点。我们将使用比色法、细胞学和免疫组织化学的刺激分析来确定这些组合的效果。我们最后的努力将是通过T细胞扩增分析来确定免疫方向。该项目风险很大，但如果成功，既可以提高研究人员对免疫系统刺激的理解，又可以创造一种指导免疫反应的方法，以加强许多潜在的疫苗。

DESCRIPTION (provided by applicant): This project aims to develop new methods to stimulate dendritic cells by finding synergistic combinations of molecular agonists. The long-term goal is to improve vaccine performance using a chemical strategy of directing the immune system. Today, vaccines are still the most effective form of disease prevention. Vaccines for many diseases remain tantalizingly close to reality, but lack high enough effectiveness to be deployed. Until recently, rationally improving vaccines was considered impossible, as vaccine creation was largely an empirical process. The cellular pathway of vaccine activation, dendritic cells, have now been identified. These cells create immunity to vaccines using a series of receptors that are stimulated by molecular agonists carried in foreign pathogens. The most effective vaccines all stimulate these receptors in synergistic combinations. Using a selected combination of molecular agonists, some vaccines direct dendritic cells to elicit either a cytotoxi immune response or a sustained, humoral response. The placement and concentration of these molecular agonists on the most effective vaccines is the critical element in creating a strong immune response. Many weaker vaccines do not use these synergistic effects. Methods for determining synergies among agonists could potentially improve weak vaccines and reveal fundamental knowledge about immune system activation. Currently however, there are no chemical methods to manipulate these molecular signals, understand their cooperative effects or attach them to potential vaccines to enhance immunity. We propose to develop a method of combining molecular agonists on inert polymeric scaffolds using a series of bio-conjugation reactions. We will use these combinations of agonists to study and understand their synergistic activity in stimulating dendritic cells. Our hypothesis is that facilitated by our scaffold, differnt combinations of agonist will direct the immune system toward greater humoral or cytotoxic immunity. We will also develop methods of coupling our immune-directing polymers onto developed vaccines, such as Enterotoxigenic E. Coli, that were not effective enough for commercial deployment. We will use hetero-telechelic polymer synthesis and an array of bio- conjugation reactions to create a combinatorial library of agonists that target many of the Toll-like and NOD-like receptors on dendritic cells responsible for immune stimulation. We will determine the effect of these combinations using colorimetric, cellular, and immunohistochemical assays of stimulation. Our final effort will be in determining immune direction via T-cell expansion assays. The project is risky, but if successful, could both improve researchers understanding of the stimulation of the immune system and create a method of directing the immune response for strengthening many potential vaccines.