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

在公众和科学界，人们普遍认为压力和免疫力之间存在着某种关系。生物过程，如免疫和压力，不存在于封闭系统的功能隔离，而是共享相当大的功能重叠和多方向的影响。这些影响通常表现在细胞信号传导水平上，以影响最终的表型。这项提案的目的是获得先天免疫信号的详细图片，并定义在急性应激的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影响先天免疫的分子机制，从而导致对感染性疾病的易感性。我们将进行激酶组分析，以精确地定义与先天免疫受体（Toll样受体）的中央家族的激活相关的信号事件，并随后调查这些信号模式的变化在应激诱导的（皮质醇和儿茶酚胺）反应的生物背景。所采用的肽阵列技术提供了前所未有的机会来破译复杂的信号转导事件，包括鉴定调节点、信号传导中间体和单一串扰。作为应激相关应答激活的结果，先天免疫信号传导的改变可能包括信号传导应答的性质（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）开发牛特异性肽激酶组阵列，其被定制为优先考虑与TLR和应激相关信号传导相关的信号传导事件，B）精确描述与Toll样受体家族不同成员的激活相关的信号传导事件，以及c）定义应激背景下这些信号传导模式的变化。这种对先天免疫信号传导（包括压力的影响）的精细视角对于理解和治疗炎症和感染（包括通过改善对动物压力的识别和管理来限制牲畜中的传染病）具有免疫学意义和实际重要性。这是至关重要的贡献，因为传染病是一个面临越来越多的抗生素使用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观点来看，通过识别和尽量减少造成压力的管理措施或选择具有更大压力耐受性的动物来降低牲畜中传染病的发病率和严重程度是非常理想的结果。

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.