An antiserum against Atlantic salmon IFNa1 detects IFN and neutralizes antiviral activity produced by poly I:C stimulated cells
An antiserum against Atlantic salmon IFNa1 detects IFN and neutralizes antiviral activity produced by poly I:C stimulated cells
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DOI：
10.1016/j.dci.2008.11.005
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发表时间：
2009-01-01
影响因子：
2.9
通讯作者：
Robertson, Borre
中科院分区：
文献类型：
--
作者：
Berg, Kristel;Svingerud, Tina;Robertson, Borre
I型干扰素（ifn）在脊椎动物抗病毒感染的先天免疫应答中起着至关重要的作用。两个ifn （IFNa1和IFNa2）此前已经从大西洋鲑鱼中克隆出来。本研究采用免疫印迹法对三文鱼IFNa1产生多克隆抗血清，并对其抗病毒活性进行中和。该抗血清首次被证实能够检测并中和HEK293细胞中产生的重组鲑鱼IFNa1的抗病毒活性。该抗血清还检测到IFNa1，并在poly I:C刺激的三文鱼TO细胞上清液中中和95-98%的抗病毒活性。这表明IFNa1/IFNa2是poly I:C刺激TO细胞产生的主要ifn。抗血清中和了5个个体中3个受poly I:C刺激的头肾白细胞的大部分IFN活性，但在其他2个个体的受刺激的白细胞中，只有75%的抗病毒活性被中和。这表明，虽然IFNa1/IFNa2是由poly I:C刺激的白细胞分泌的主要ifn，但这些细胞也可以产生具有ifn样活性的其他分子。(C) 2008 Elsevier Ltd版权所有。
Type I interferons (IFNs) play a crucial role in innate immune responses against virus infections in vertebrates. Two IFNs (IFNa1 and IFNa2) have previously been cloned from Atlantic salmon. In the present work a polyclonal antiserum, which was generated against salmon IFNa1 was used to study its production in cells by immunoblot detection and neutralization of antiviral activity. The antiserum was first confirmed to detect and neutralize the antiviral activity of recombinant salmon IFNa1 produced in HEK293 cells. The antiserum also detected IFNa1 and neutralized 95-98% of the antiviral activity in supernatants of poly I:C stimulated salmon TO cells. This suggests that IFNa1/IFNa2 are the major IFNs produced by poly I:C stimulated TO cells. The antiserum neutralized most of the IFN activity in poly I:C stimulated head kidney leucocytes from three of five individuals, but in stimulated leucocytes from the other two individuals only 75% of the antiviral activity was neutralized. This shows that although IFNa1/IFNa2 are major IFNs secreted by poly I:C stimulated leucocytes, these cells can also produce additional molecules with IFN-like activity. (C) 2008 Elsevier Ltd. All rights reserved.