Characterization of Crustacean Innate Immune Response

对动物免疫学的广泛理解需要了解来自不同进化谱系的物种中特定免疫相关分子的多样性。先天免疫反应是脊椎动物和无脊椎动物共有的基本免疫过程，这一点尤其正确。然而，我们目前对先天免疫的理解主要集中在模式生物上，如人类和小鼠；那里有大量的基因组资源。我们对非模式物种先天免疫反应的广度和范围的理解受到我们无法使用高通量基因组方法的限制。基因组测序技术的最新进展允许在非模式生物中全面检查和发现免疫相关转录物。新的免疫分子的发现将需要病原体挑战实验、差异表达和生物信息学分析以及定向功能分析来全面确定生物功能。节肢动物门是地球上数量最多、生物学上最成功的动物；海洋被各种甲壳类动物所主宰。无脊椎动物的比较先天免疫学主要集中在陆生无脊椎动物，如果蝇，这给我们对甲壳类动物免疫学的认识留下了很大的空白。甲壳类动物免疫学的传统方法依赖于光和电子显微镜方法。高通量测序方法的应用极大地扩展了我们对无脊椎动物免疫学的理解。我最近在病原体和环境胁迫下生成了美国龙虾（Homarus americanus）的多个转录组；以及与美洲人共享栖息地的几种甲壳类动物的转录组。这些转录组资源为在生物信息学和功能水平上研究比较免疫学提供了令人兴奋的基础。我的研究项目的长期目标是了解甲壳类动物先天免疫的能力和机制。我的建议有三个主要目标：目标1。探讨十足甲壳类动物模式识别受体、toll样受体、免疫缺陷细胞内信号分子和抗菌肽的保守性和多样性。目标2。发现美洲龙虾（Homarus americanus）对细菌感染反应相关基因并进行功能注释。目标3。表征抗菌肽对海洋甲壳类动物特定致病菌和条件致病菌的生物活性。这些研究将拓宽我们对美洲Homarus americanus先天免疫的生理机制的理解，以及更广泛的无脊椎动物免疫学。

A broad understanding of animal immunology requires understanding the diversity of specialized immune-related molecules in species from diverse evolutionary linages. This is especially true for innate immune responses which are fundamental immunological processes shared by vertebrate and invertebrate animals. However, our current understanding of innate immunity is focused on model organisms such as humans and mice; where extensive genomic resources exist. Our understanding of the breadth and scope of innate immune responses in non-model species has been limited by our inability to use high-through genomic approaches. Recent advances in genomic sequencing technology permit the comprehensive examination and discovery of immune-related transcripts in non-model organisms. Novel immune molecule discovery will require pathogen challenge experiments accompanied by differential expression and bioinformatic analysis and directed functional assays to comprehensively assign biological function.The Arthropoda phylum encompasses the most abundant and biologically successful animals on earth; with the oceans dominated by diverse species of crustaceans. Comparative innate immunology in invertebrates has largely focused on terrestrial invertebrates such as Drosophila spp. This has left major gaps in our knowledge of crustacean immunology. Traditional approaches to crustacean immunology relied on light and electron microscopy approaches. The application of high-throughput sequencing approaches holds great promise for significantly expanding our understanding of invertebrate immunology.I have recently generated multiple transcriptomes of the American lobster (Homarus americanus) under pathogen and environmental stress challenges; as well as transcriptomes from several crustacean species which share habitats with H. americanus. These transcriptomic resources represent an exciting basis for examining comparative immunology at both the bioinformatic and functional level. The long-term goals of my research program focus on understanding the capabilities and mechanisms underlying crustacean innate immunity. I have three major objectives for this proposal:Objective 1. To determine the conservation and diversity of pattern recognition receptors, Toll-like receptor and Immunodeficiency intracellular signaling molecules and antimicrobial peptides among decapod crustaceans. Objective 2. To discover and functionally annotate American lobster (Homarus americanus) genes involved in responding to bacterial infection. Objective 3. To characterize the bioactivity of antimicrobial peptides against a panel of specific and opportunistic pathogens of marine crustaceans.These studies will broaden our understanding of the physiological mechanisms underlying Homarus americanus innate immunity specifically, and invertebrate immunology more broadly.