Kileng Øyvind, Brundtland Marthe Iren, Robertsen Børre
Department of Marine Biotechnology, Norwegian College of Fishery Science, University of Tromsø, Breivika, N-9037 Tromsø, Norway.
Fish Shellfish Immunol. 2007 Aug;23(2):378-89. doi: 10.1016/j.fsi.2006.11.011. Epub 2006 Dec 3.
Infectious salmon anemia virus (ISAV) is an aquatic orthomyxovirus causing disease and high mortality in farmed Atlantic salmon (Salmo salar). The virus is thus apparently able to initiate replication without being hampered by the host's immune system. In this work we have studied the role of the type I interferon (IFN) system of Atlantic salmon in protection against ISAV. Real-time RT-PCR was used to study the expression of type I IFN and the IFN stimulated genes Mx and ISG15 in TO cells and live fish in response to infection with ISAV. The in vitro studies showed that ISAV was a powerful inducer of Mx and ISG15 genes in TO cells and that induction started relatively early during infection. In contrast, IFN transcripts were induced later than both Mx and ISG15 transcripts in the ISAV infected cells indicating that Mx and ISG15 are induced through IFN-independent pathways in the early stages of ISAV infection. A cohabitee infection trial with ISAV in Atlantic salmon resulted in high mortality, even though elevated levels of IFN, Mx and ISG15 transcripts in the head kidney and liver were observed. Immunoblotting confirmed the presence of Mx and ISG15 proteins in the liver of infected salmon. In order to evaluate whether the type I IFN system is able to inhibit replication of ISAV, TO cells were stimulated with recombinant salmon IFN-alpha1 (rSasaIFN-alpha1) and subsequently infected with virus. The rSasaIFN-alpha1 showed no protection of TO cells against ISAV, but full protection against IPNV. These data demonstrate that key proteins of the type I IFN system are induced during an ISAV infection, but that they are unable to inhibit the replication of ISAV in vitro and in vivo. ISAV must thus encode genes that enable the virus to counteract IFN induced antiviral proteins of the host.
传染性鲑鱼贫血病毒(ISAV)是一种水生正粘病毒,可导致养殖大西洋鲑(Salmo salar)患病并造成高死亡率。因此,该病毒显然能够在不受宿主免疫系统阻碍的情况下启动复制。在本研究中,我们探究了大西洋鲑的I型干扰素(IFN)系统在抵御ISAV中的作用。利用实时逆转录聚合酶链反应(RT-PCR)研究了I型IFN以及IFN刺激基因Mx和ISG15在TO细胞和活体鱼中对ISAV感染的反应表达情况。体外研究表明,ISAV是TO细胞中Mx和ISG15基因的强力诱导剂,且诱导在感染过程中相对较早开始。相比之下,在ISAV感染的细胞中,IFN转录本的诱导晚于Mx和ISG15转录本,这表明在ISAV感染早期,Mx和ISG15是通过不依赖IFN的途径被诱导的。在大西洋鲑中进行的ISAV同居感染试验导致了高死亡率,尽管在头肾和肝脏中观察到IFN、Mx和ISG15转录本水平升高。免疫印迹证实了感染鲑鱼肝脏中存在Mx和ISG15蛋白。为了评估I型IFN系统是否能够抑制ISAV的复制,用重组鲑鱼IFN-α1(rSasaIFN-α1)刺激TO细胞,随后用病毒感染。rSasaIFN-α1对TO细胞没有表现出对ISAV的保护作用,但对传染性胰腺坏死病毒(IPNV)有完全的保护作用。这些数据表明,I型IFN系统的关键蛋白在ISAV感染期间被诱导,但它们在体外和体内均无法抑制ISAV的复制。因此,ISAV必须编码能够使该病毒对抗宿主IFN诱导的抗病毒蛋白的基因。