Harnessing interleukin 2 (IL-2) family power in fish; study of rainbow trout IL-2, IL-15, IL-15L and IL-15Ralpha

利用鱼类中白细胞介素 2 (IL-2) 家族的力量；

• 批准号: 317850006
• 负责人: Privatdozent Dr. Uwe Fischer
• 金额: --
• 依托单位: Institut für Infektionsmedizin (IMED)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2016
• 资助国家: 德国
• 起止时间: 2015-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/317850006?language=en
• 关键词: Harnessing; interleukin; IL; family; power

Interleukins (ILs) are a group of cytokines that control many immune functions. In mammals, IL-2 and IL-15 are potent growth factors for cytotoxic, T helper and B cells. These cytokines are also involved in the formation and maintenance of immunological memory. Fishes, like other vertebrates, possess an immunological memory and thus can be vaccinated. In mammals, proper function of IL-15 is ensured in association with the IL-15 receptor (R) alpha chain. The function of another related cytokine, IL-15 like (IL-15L), remains unclear even in higher vertebrates. The original project was supported by the DFG to study the effect of IL-2 family cytokines in association with IL-15Ralpha in rainbow trout. The main conclusions from this project were: - fusion of IL-15 and IL-15L to sIL-15Ralpha (RLI) enhances cytokine secretion;- IL-15-RLI efficiently stimulates cytotoxic NK cells and lymphocytes from mucosal organs;- main IL-15L-RLI targets CD4-CD8-IgM- cells;- IL-2 family cytokines differently stimulate the developmental stages of trout thymocytes.However, our most fundamental discovery was that IL-15 triggers type 1 and that, in contrast, IL-15L induces type 2 immune gene transcription. Now, the functional relevance of these latter observations remains to be shown in vivo. Thus, in the project renewal, I apply for finalizing our work on in vivo immune responses behind these type 1 and type 2 triggers: cell-mediated cytotoxicity and antibody responses, respectively. Special attention will be given to the capacity of trout IL-2 family cytokines to extend the immunological memory after vaccination to overcome the short term-protection of current fish vaccines. These studies will be backed by whole immune transcriptome analysis and provide complex insights into immune pathway regulation. To address this final goal we have available six essential toolboxes:- clonal trout, a major aquacultured species, as model organism;- established platform for cytokine production;- trout leukocyte markers;- functional assays;- infection models;- whole transcriptome platform.This toolbox enables us to pursue the following main objectives: (1) in vitro cytotoxic activity of leukocytes isolated from cytokine-treated fish and (2) in vivo effects of cytokines on immune cell dynamics in naïve fish; on the outcome of viral and bacterial infections; on the effects on long-term protection and immune memory.The project renewal application aims at a duration of 24 months. The main justifications for this duration is the time needed to study immune memory since decline of vaccine-induced protection usually starts approx.12 months after immunization. Considering the time needed to produce recombinant cytokines, to get the approval for animal experiments from the ethics commission, to vaccinate and challenge (viral and bacterial) the fish and to acquire and analyse the data, I believe that a project duration of 24 months is justifiable.

白细胞介素（IL）是一组控制多种免疫功能的细胞因子。在哺乳动物中，IL-2和IL-15是细胞毒性T辅助细胞和B细胞的有效生长因子。这些细胞因子也参与免疫记忆的形成和维持。鱼类和其他脊椎动物一样，具有免疫记忆，因此可以接种疫苗。在哺乳动物中，IL-15的适当功能与IL-15受体（R）α链相关。另一种相关细胞因子IL-15样（IL-15 L）的功能即使在高等脊椎动物中也仍不清楚。最初的项目是由DFG支持的，以研究IL-2家族细胞因子与虹鳟鱼中IL-15 R α相关的作用。该项目的主要结论是：-IL-15和IL-15 L与sIL-15 R α（RLI）的融合增强细胞因子分泌;- IL-15-RLI有效地刺激来自粘膜器官的细胞毒性NK细胞和淋巴细胞;-主要的IL-15 L-RLI靶向CD 4-CD 8-IgM-细胞;- IL-2家族细胞因子不同地刺激鳟鱼胸腺细胞的发育阶段。我们最基本的发现是IL-15触发1型免疫基因转录，相反，IL-15 L诱导2型免疫基因转录。现在，这些后一种观察结果的功能相关性仍有待于在体内显示。因此，在项目更新中，我申请完成我们在这些1型和2型触发因素背后的体内免疫反应方面的工作：分别是细胞介导的细胞毒性和抗体反应。将特别关注鳟鱼IL-2家族细胞因子在疫苗接种后延长免疫记忆的能力，以克服目前鱼类疫苗的短期保护。这些研究将得到全免疫转录组分析的支持，并为免疫途径调控提供复杂的见解。为了实现这一最终目标，我们提供了六个基本工具箱：-克隆鳟鱼，一种主要的水产养殖物种，作为模式生物;-建立细胞因子生产平台;-鳟鱼白细胞标志物;-功能测定;-感染模型;-全转录组平台。这个工具箱使我们能够实现以下主要目标：（1）从经精氨酸处理的鱼中分离的白细胞的体外细胞毒性活性和（2）细胞因子对幼稚鱼中免疫细胞动力学的体内影响;病毒和细菌感染的结果;对长期保护和免疫记忆的影响。项目续期申请旨在为期24个月。这一持续时间的主要理由是研究免疫记忆所需的时间，因为疫苗诱导的保护作用通常在免疫后约12个月开始下降。考虑到生产重组细胞因子、获得伦理委员会对动物实验的批准、对鱼进行疫苗接种和挑战（病毒和细菌）以及获取和分析数据所需的时间，我认为24个月的项目期限是合理的。