Small molecule agonists and antagonists of inflammatory responses mediated by Toll- and Nod- like receptors

人体不断暴露于细菌和病毒等微生物。其中一些与人类生活在一个快乐的平衡中，而另一些则会损害身体并导致疾病。后一组被称为病原体。人类通过一个非常复杂但经过精细调整的系统来识别病原体并做出反应，称为免疫反应。这是由两种不同但互补的成分形成的，称为适应性免疫反应和先天免疫反应。适应性免疫反应是针对特定病原体的，需要几天时间才能完全激活，并且是疫苗引发的反应类型。先天反应是即时的，对所有病原体的反应都是一样的。它在感染附近创造了一种抗病原体的状态，在这种状态下，细胞准备好对抗病原体。通过先天免疫反应检测微生物是极其敏感的。它使用细胞表面和细胞内部的蛋白质来识别被称为PAMP（病原体相关分子模式）的微生物的小部分。PAMP对于微生物功能是必需的，并且在微生物之间显示很少或没有变化。PAMP可以包括细菌细胞壁的组分，或病毒和细菌的核酸基因组中的序列。通过更好地了解PAMP的检测和相关过程，科学家有可能在未来的感染治疗中产生深远的影响。我们的研究将调查两个家族的人类蛋白质参与识别PAMP和激活先天免疫反应的功能。它们是Toll样受体（TLR）和NOD样受体（NLR）。特别是，我们将专注于称为TLR 8，NOD 1和NALP 1的单个蛋白质。这些细胞识别来自病毒的遗传物质和细菌周围的部分细胞壁。通过了解这些蛋白质的功能，我们将能够改善感染和其他疾病，如类风湿性关节炎，炎症性肠病和哮喘的治疗方法。事实上，一些已经用于治疗病毒和肿瘤的药物含有与TLR 8相互作用并帮助其发挥作用的小分子。为了了解这些蛋白质是如何工作的，我们需要知道它们在细胞中是如何打开和关闭的，以及它们如何与药物分子和PAMP相互作用。蛋白质形成的形状对这些过程非常重要。在我们的研究中，我们将在细菌和病毒中培养这些蛋白质的不同部分。然后这些将被净化，以允许我们与他们一起工作。我们将研究这些蛋白质如何与药物和其他可以制成药物的小分子结合。这将使用技术，如表面等离子体共振和等温滴定量热法。这些技术使我们能够将蛋白质和药物混合，以观察它们相互结合的速度和强度。我们还将寻找与蛋白质结合的新分子，看看它们是否使蛋白质更活跃或更不活跃。最后，我们将生长蛋白质晶体，让我们用一种叫做X射线晶体学的技术来观察它们的形状。这可以通过观察蛋白质晶体的X射线照片形成的图案来提供蛋白质三维形状的线索。这些图片将告诉我们蛋白质如何与其他分子相互作用。这项工作将解释TLR 8，NOD 1和NALP 1如何对抗感染;它们如何对药物作出反应;以及它们的活性如何改变。特别是，它还将确定未来可用作药物的新分子，以改善对感染和炎症性疾病的治疗。这些将有可能对全球一级的卫生管理产生重大的积极影响。

The human body is continually exposed to micro-organisms such as bacteria and viruses. Some of these live in a happy balance with humans, whilst others can damage the body and cause illness and disease. This latter group are known as pathogens. Humans recognise and respond to pathogens through an amazingly complex, but finely tuned system known as the immune response. This is formed from two different, but complementary, components called the adaptive and innate immune responses. The adaptive immune response is specific to a particular pathogen, takes a few days to become fully active and is the type of response primed by vaccines. The innate response is immediate, and responds in the same way to all pathogens. It creates an anti-pathogen state near the infection in which cells are primed to fight the pathogens. The detection of micro-organisms by the innate immune response is exquisitely sensitive. It uses proteins both on the surface and inside cells to recognise small parts of micro-organisms known as PAMPs (Pathogen Associated Molecular Patterns). PAMPs are essential for micro-organism function and show little or no variation between micro-organisms. PAMPs may include components of the bacterial cell wall, or sequences in the nucleic acid genome of viruses and bacteria. Through a better understanding of the detection of PAMPs and associated processes scientists have the potential to make far-reaching impacts in the treatment of infection in the future. Our research will investigate the function of two families of human proteins involved in recognising PAMPs and activating the innate immune response. These are the Toll-like receptors (TLRs) and the NOD-like receptors (NLRs). In particular we will focus on individual proteins called TLR8, NOD1 and NALP1. These recognise bits of genetic material from viruses and parts of the cell wall that surround bacteria. By understanding how these proteins function we will be able to improve the ways that infections and other diseases such as rheumatoid arthritis, inflammatory bowel disease and asthma can be treated. Indeed some of the drugs already available to treat viruses and tumours contain small molecules that interact with TLR8 and help it work. In order to understand how these proteins work we need to know how they are switched on and off in a cell and how they interact with drug molecules and PAMPs. The shape that the proteins make is very important for these processes. In our research we will grow different parts of these proteins in bacteria and viruses. These will then be purified to allow us to work with them. We will investigate how these proteins bind to drugs and other small molecules that could be made into drugs. This will use techniques such as surface plasmon resonance and isothermal titration calorimetry. These techniques allow us to mix the protein and drug to see how quickly and strongly they bind to each other. We will also look for new molecules that bind to the proteins and see whether they make the proteins more or less active. Finally we shall grow crystals of the proteins to let us look at their shape using a technique called X-ray crystallography. This can give clues to a protein's 3-dimensional shape by looking at the patterns formed by taking X-ray pictures of protein crystals. These pictures will tell us exactly how the proteins interact with other molecules. This work will explain how TLR8, NOD1 and NALP1 work to fight infection; how they react to drugs; and how their activity can be changed. In particular it will also identify new molecules that could be used as drugs in the future to improve treatments against infection and inflammatory diseases. These would have the potential to make a significant positive impact to health management on a global level.