Novel Immune-Type Receptors and Cytotoxicity In Zebrafish

斑马鱼的新型免疫型受体和细胞毒性

• 批准号: 0344618
• 负责人: Jeffrey Yoder
• 金额: $ 3.4万
• 依托单位: University of South Florida
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2005-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0344618&HistoricalAwards=false
• 关键词: Novel; Immune; Type; Receptors; Cytotoxicity

PROJECT SUMMARYIn many vertebrates, one function of the innate immune system is carried out by cytotoxic cells that recognize and destroy virally infected or cancerous cells. The molecular mechanisms involved in these processes are well studied in mammalian natural killer (NK) cells, but very little is known about them in other species. A family of Novel Immune Type Receptor (NITR) genes has recently been identified in bony fish with characteristics similar to the mammalian NK receptors. Thus, it has been hypothesized that these genes play a role in cytotoxic cell function in fish. This project will determine whether zebrafish NITR gene families are expressed in cell lineages known to possess cytotoxic properties and whether zebrafish NITR-expressing cells exhibit cytotoxic activity. First, tissue-specific expression patterns will be determined for multiple NITR gene families by RT-PCR and RNA blot analysis. Second, expression of NITR genes will be examined by RNA in situ hybridization in cell lineages known to be cytotoxic. Third, transgenic lines of zebrafish will be generated and analyzed in which the Green Fluorescent Protein will serve as an in vivo fluorescent biomarker for NITR-expressing cells, which will then be isolated and tested for cytotoxic activity. Knowledge gained from this project will provide new insight into the function of cytotoxic signaling pathways and elucidate the molecular mechanisms of innate immune reactivity in fish.The results of this project will contribute to a better understanding of immune function in fish, including those of agricultural importance and will have implications for understanding the molecular mechanisms of cytotoxic defense in organisms that occupy vastly different environmental niches and encounter diverse pathogens. This project will involve multiple graduate and undergraduate students who will gain expertise in various genetic, molecular and biochemical techniques, while being trained in experimental design and the scientific process.

在许多脊椎动物中，天然免疫系统的一项功能是由识别和摧毁病毒感染或癌细胞的细胞毒细胞执行的。参与这些过程的分子机制在哺乳动物自然杀伤(NK)细胞中得到了很好的研究，但在其他物种中却知之甚少。最近在硬骨鱼中发现了一个新的免疫类型受体(NITR)基因家族，其特征与哺乳动物的NK受体相似。因此，人们假设这些基因在鱼类的细胞毒细胞功能中发挥作用。该项目将确定斑马鱼NITR基因家族是否在已知具有细胞毒活性的细胞系中表达，以及表达斑马鱼NITR的细胞是否具有细胞毒活性。首先，通过RT-PCR和RNA印迹分析确定多个NITR基因家族的组织特异性表达模式。其次，NITR基因的表达将通过RNA原位杂交在已知具有细胞毒性的细胞系中进行检测。第三，将产生并分析斑马鱼的转基因株系，其中绿色荧光蛋白将作为表达NITR的细胞的体内荧光生物标记物，然后将其分离并检测其细胞毒活性。本项目所获得的知识将对细胞毒信号通路的功能提供新的见解，并阐明鱼类先天免疫反应的分子机制。本项目的结果将有助于更好地了解鱼类的免疫功能，包括那些具有农业重要性的免疫功能，并将对理解生物中细胞毒防御的分子机制具有重要意义，这些生物所处的环境生态位差异很大，并面临着不同的病原体。该项目将涉及多名研究生和本科生，他们将获得各种遗传、分子和生化技术方面的专业知识，同时接受实验设计和科学过程方面的培训。