Kinomic Evaluation of Toll-like Receptor Signalling: Specialization, Synergies and Stress

在普通公众和科学界中，普遍认识到压力和免疫力之间存在着关系。生物过程，如免疫和应激，并不存在于封闭系统的功能隔离中，而是存在相当大的功能重叠和多方向影响。这些影响通常表现在细胞信号水平上，以影响最终的表型。这项建议的目的是获得先天免疫信号的详细图景，并确定在急性应激背景下这些模式的变化。最终目标是确定应激反应的不同分支通过影响先天免疫从而导致传染病易感性的分子机制。我们将进行动态组分析，以精确定义与天然免疫受体(Toll样受体)中央家族激活相关的信号事件，并随后在应激诱导(皮质醇和儿茶酚胺)反应的生物学背景下研究这些信号模式的变化。将采用的多肽阵列技术为破译复杂的信号转导事件提供了前所未有的机会，包括识别调节点、信号传递中间体和分离串扰。作为应激相关反应激活的结果，先天免疫信号的改变可能包括信号反应的性质(途径、蛋白质、磷酸化事件)、幅度或持续时间，并且可能以特定TLR特有的方式发生。目前研究的重点是：a)开发牛特异性多肽激动组阵列，该阵列是定制的，用于优先处理与TLR和应激相关的信号事件；b)精确描述与Toll样受体家族不同成员激活相关的信号事件；c)确定这些信号模式在应激背景下的变化。这种关于先天性免疫信号的精细视角，包括应激的影响，对于理解和治疗炎症和感染，包括通过改进对动物应激的认识和管理来限制家畜的传染病，将具有免疫学上的兴趣和实际意义。这是至关重要的贡献，因为传染病是面临抗生素使用越来越多限制的行业的最大经济损失来源。从动物健康和福利以及行业认知的角度来看，通过确定和最大限度地减少造成压力的管理做法或选择具有更高压力耐受性的动物来降低牲畜传染病的发生率和严重程度是非常可取的结果。

Within both the general public and scientific community there is an overall appreciation that a relationship exists between stress and immunity. Biological processes, such as immunity and stress, do not exist in the functional isolation of closed systems but rather share considerable functional overlap and multidirectional influence. The influences often manifest at the level of cellular signalling to influence the ultimate phenotypes. The objective of this proposal is to obtain a detailed picture of innate immune signaling and to define changes to those patterns within the context of acute stress. The ultimate goal is to to define molecular mechanisms by which the different branches of the stress response influence innate immunity with subsequent consequences to susceptibility to infectious disease. We will conduct kinome analysis to precisely define signaling events associated with activation of a central family of innate immune receptors (Toll-like receptors) and subsequently investigate changes to these signalling patterns within the biological context of stress-induced (cortisol and catecholamine) responses. The peptide arrays technology to be employed offers unprecedented opportunity to decipher complex signal transduction events, including identification of points of regulation, signalling intermediates and singling cross-talk. Alterations to innate immune signalling as a consequence of activation of stress-associated responses may include the nature (pathways, proteins, phosphorylation events), magnitude, or duration of signalling responses and may occur in a manner that is unique to specific TLRs. The priorities of the current investigation are to: a) develop a bovine-specific peptide kinome array that is customized to prioritize signalling events associated with TLR and stress-associated signaling, b) precisely describe signaling events associated with activation of different members of the Toll-like receptor family, and c) define changes to these signaling patterns in the context of stress. This refined perspective on innate immune signaling, including the influences of stress, will be of both immunological interest and practical importance for understanding and treating inflammation and infections, including limiting infectious disease in livestock through improved recognition and management of animal stress. This is critical contribution as infectious diseases represent the greatest source of economic losses to an industry facing increasing restrictions on the use of antibiotics. Reducing the incidence and severity of infectious diseases in livestock by identifying and minimizing management practices that cause stress, or selecting animals with greater stress tolerance, is a highly desirable outcome from the perspectives of animal health and welfare, as well as industry perception.