Maehr Tanja, Vecino Jose L González, Wadsworth Simon, Wang Tiehui, Secombes Christopher J
Scottish Fish Immunology Research Centre, Institute of Biological and Environmental Sciences, University of Aberdeen, Aberdeen AB24 2TZ, UK.
EWOS Innovation AS, 4335 Dirdal, Norway.
Mol Immunol. 2014 Nov;62(1):186-98. doi: 10.1016/j.molimm.2014.06.021. Epub 2014 Jul 9.
Suppressor of cytokine signalling (SOCS) family members are crucial in the control and attenuation of cytokine induced responses via activation of the JAK/STAT, TLR and NF-kB signalling pathways. SOCS proteins orchestrate the termination of many types of immune responses and are often the targets of microbial pathogens exploiting SOCS mechanisms to evade the host's immune response. Through whole and lineage specific genome duplication events, the teleost cytokine/SOCS network is complex. Not only are the orthologues of all mammalian SOCS members present, namely cytokine inducible Src homology 2 (SH2)-containing protein (CISH) and SOCS-1 to -7, but multiple gene copies exist that may potentially become functionally divergent. In this paper we focus on the CISH genes in rainbow trout (Oncorhynchus mykiss), and have cloned two further paralogues, CISHa2 and CISHb2, additional to the known CISHa1 and CISHb1 genes. We present for the first time a comparative expression analysis of these four paralogues, to establish whether subfunctionalisation is apparent. In vivo examination of gene expression revealed a higher constitutive expression level of CISHa paralogues compared to CISHb expression in adult trout tissues. All CISHs were relatively highly abundant in immune tissues but CISHa2 and CISHb2 had highest expression in the heart and muscle. An inverse picture of CISH abundance during trout ontogeny was seen, and further hints at differential roles of the four genes in immune regulation and development. Stimulation of head kidney (HK) leukocytes with trout recombinant interleukin (rIL)-15 and rIL-21 had a major effect on CISHa2 and to a lesser extent CISHa1 expression. In HK macrophages rIL-1β, phytohemagglutinin, and phorbol 12-myristate 13-acetate also had a strong impact on CISHa2 expression. Yersinia ruckeri infection caused a temporally and spatially differential onset of CISH expression that may be viewed in the context of pathogen evasion strategies. These data, against the backdrop of fish specific whole genome duplication events and functional divergence, provide the first evidence for differential roles of the four trout CISH genes in immune control and development.
细胞因子信号转导抑制因子（SOCS）家族成员在通过激活JAK/STAT、TLR和NF-κB信号通路来控制和减弱细胞因子诱导的反应中起着关键作用。SOCS蛋白协调多种免疫反应的终止，并且常常成为微生物病原体利用SOCS机制逃避宿主免疫反应的靶点。通过全基因组和谱系特异性基因组复制事件，硬骨鱼的细胞因子/SOCS网络很复杂。不仅存在所有哺乳动物SOCS成员的直系同源物，即细胞因子诱导含Src同源2（SH2）结构域蛋白（CISH）以及SOCS-1至-7，而且还存在多个可能在功能上发生分化的基因拷贝。在本文中，我们聚焦于虹鳟（Oncorhynchus mykiss）的CISH基因，除了已知的CISHa1和CISHb1基因外，又克隆了另外两个旁系同源物CISHa2和CISHb2。我们首次对这四个旁系同源物进行了比较表达分析，以确定是否存在明显的亚功能化现象。对基因表达的体内检测显示，在成年鳟鱼组织中，CISHa旁系同源物的组成型表达水平高于CISHb。所有CISH在免疫组织中相对丰度较高，但CISHa2和CISHb2在心脏和肌肉中表达最高。在鳟鱼个体发育过程中观察到CISH丰度的相反情况，这进一步暗示了这四个基因在免疫调节和发育中的不同作用。用鳟鱼重组白细胞介素（rIL）-15和rIL-21刺激头肾（HK）白细胞对CISHa2有主要影响，对CISHa1的影响较小。在HK巨噬细胞中，rIL-1β、植物血凝素和佛波酯12-肉豆蔻酸酯13-乙酸酯也对CISHa2表达有强烈影响。鲁氏耶尔森菌感染导致CISH表达在时间和空间上出现差异，这可能与病原体的逃避策略有关。这些数据，在鱼类特异性全基因组复制事件和功能分化的背景下，为四个鳟鱼CISH基因在免疫控制和发育中的不同作用提供了首个证据。
