虾源性转座酶衍生 1（HARBI1）样分子特征及其在白斑综合征病毒和溶藻弧菌感染中的作用。
Molecular characterization of shrimp harbinger transposase derived 1 (HARBI1)-like and its role in white spot syndrome virus and Vibrio alginolyticus infection.
机构信息
College of Animal Science and Technology, Zhejiang Agriculture and Forestry University, Hangzhou 311300, China.
College of Animal Science and Technology, Zhejiang Agriculture and Forestry University, Hangzhou 311300, China.
出版信息
Fish Shellfish Immunol. 2018 Jul;78:222-232. doi: 10.1016/j.fsi.2018.04.032. Epub 2018 Apr 19.
The role of the nuclease, HARBI1-like protein (mjHARBI1-like) in the innate immunity of Marsupenaeus japonicus was explored in this study. The 1361 bp cDNA sequence of mjHARBI1-like was cloned from M. japonicus using RACE. RT-qPCR analysis results showed that the gills and hepatopancreas of M. japonicus were the main tissues where mjHARBI1-like is expressed. In addition, it was also found that white spot syndrome virus (WSSV) or Vibrio alginolyticus challenge could stimulate mjHARBI1-like expression. After mjHARBI1-likewas inhibited, expression of immune genes such as toll, p53, myosin, and proPO were significantly downregulated (P < 0.01). However, in shrimp hemocytes, hemocyanin and tumor necrosis factor-α (TNF-α) were up-regulated significantly (P < 0.01). This study demonstrated that mjHARBI1-like plays a key role in the progression of WSSV and V. alginolyticus infection. Specifically, the cumulative mortality of WSSV-infected and V. alginolyticus-infected shrimp was significantly advanced by double-strand RNA interference (dsRNAi) of mjHARBI1-like. Apoptosis studies indicated that mjHARBI1-dsRNA treatment caused a reduction in hemocyte apoptosis in bacterial and viral groups. In addition, phagocytosis experiments illustrated that mjHARBI1-dsRNA treatment led to a lower phagocytosis rate in hemocytes of V. alginolyticus-challenged shrimp. It was also found that knockdown of mjHARBI1-like inhibited shrimp phenoloxidase (PO) activity, superoxide dismutase (SOD) activity, and total hemocyte count (THC) after WSSV or V. alginolyticus infection. These data indicate a regulative role of mjHARBI1-likein the immunity of shrimp in response to pathogen infection. Resultantly, it was concluded that mjHARBI1-like might have a positive effect on the anti-WSSV immune response of shrimp by regulating apoptosis, THC, PO activity, and SOD activity. Additionally, mjHARBI1-like might promote anti-V. alginolyticus infection by participating in regulating phagocytosis, apoptosis, SOD activity, PO activity, and THC.
本研究探讨了核酸酶、HARBI1 样蛋白（mjHARBI1-like）在日本对虾先天免疫中的作用。本研究采用 RACE 技术从日本对虾中克隆出 1361 bp 的 mjHARBI1-like cDNA 序列。RT-qPCR 分析结果表明，日本对虾的鳃和肝胰腺是 mjHARBI1-like 表达的主要组织。此外，还发现白斑综合征病毒（WSSV）或溶藻弧菌的刺激可以诱导 mjHARBI1-like 的表达。抑制 mjHARBI1-like 后，免疫基因如 toll、p53、肌球蛋白和 proPO 的表达显著下调（P<0.01）。然而，在虾血细胞中，血蓝蛋白和肿瘤坏死因子-α（TNF-α）显著上调（P<0.01）。本研究表明，mjHARBI1-like 在 WSSV 和 V. alginolyticus 感染的进展中起着关键作用。具体来说，双链 RNA 干扰（dsRNAi）抑制 mjHARBI1-like 可显著提高 WSSV 感染和 V. alginolyticus 感染虾的累计死亡率。凋亡研究表明，mjHARBI1-dsRNA 处理导致细菌和病毒组血细胞凋亡减少。此外，吞噬实验表明，mjHARBI1-dsRNA 处理导致 V. alginolyticus 攻毒虾血细胞的吞噬率降低。还发现，抑制 mjHARBI1-like 的表达可抑制 WSSV 或 V. alginolyticus 感染后虾酚氧化酶（PO）活性、超氧化物歧化酶（SOD）活性和总血细胞计数（THC）。这些数据表明 mjHARBI1-like 在虾对病原体感染的免疫反应中起调节作用。因此，得出结论，mjHARBI1-like 通过调节凋亡、THC、PO 活性和 SOD 活性，可能对虾的抗 WSSV 免疫反应产生积极影响。此外，mjHARBI1-like 可能通过参与调节吞噬作用、凋亡、SOD 活性、PO 活性和 THC 来促进抗 V. alginolyticus 感染。
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