Romero Alejandro, Figueras Antonio, Thoulouze Maria-Isabel, Bremont Michael, Novoa Beatriz
Instituto de Investigaciones Marinas, CSIC, Eduardo Cabello 6, 36208 Vigo, Spain.
Dis Aquat Organ. 2008 Jul 7;80(2):123-35. doi: 10.3354/dao01932.
Infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicaemia virus (VHSV) are rhabdoviruses that infect salmonids, producing serious economic losses. Two recombinant IHN viruses were generated by reverse genetics. For one (rIHNV GFP) the IHNV NV gene was replaced with the green fluorescent protein (GFP) gene. In the other (rIHNV-Gvhsv GFP) the G gene was also exchanged for that of VHSV. No mortalities, external signs or histological lesions were observed in experimental infections conducted with the recombinant viruses. Neither the rIHNV GFP nor rIHNV-Gvhsv GFP was detected by RT-PCR in any of the examined tissues from experimentally infected fish. In order to assess their potential as vaccines against the wild type viruses, rainbow trout were vaccinated with the recombinant viruses by intraperitoneal injection and challenged 30 d later with virulent IHNV or VHSV. The GFP viruses provided protection against both wild type viruses. None of the recombinant viruses induced antibody production, and the expression of interferon (IFNalpha4) and interferon induced genes such as Mx protein and ISG-15 was not different to that of controls. The rIHNV-Gvhsv GFP did not inhibit cellular apoptosis as it was observed in an IHNV inoculated fish cell line. These studies suggest that the recombinant rIHNV-Gvhsv GFP is a promising candidate as a live recombinant vaccine and also provides a good model to further study viral pathogenicity and the molecular basis of protection against these viral infections.
传染性造血坏死病毒(IHNV)和病毒性出血性败血症病毒(VHSV)是感染鲑科鱼类的弹状病毒,会造成严重的经济损失。通过反向遗传学产生了两种重组IHN病毒。其中一种(rIHNV GFP)的IHN病毒NV基因被绿色荧光蛋白(GFP)基因取代。另一种(rIHNV-Gvhsv GFP)的G基因也被VHSV的G基因替换。在用重组病毒进行的实验感染中未观察到死亡、外部症状或组织学损伤。在实验感染鱼的任何检测组织中,通过RT-PCR均未检测到rIHNV GFP和rIHNV-Gvhsv GFP。为了评估它们作为针对野生型病毒的疫苗的潜力,虹鳟鱼通过腹腔注射接种重组病毒,并在30天后用强毒IHNV或VHSV进行攻毒。GFP病毒对两种野生型病毒均提供了保护。重组病毒均未诱导抗体产生,并且干扰素(IFNalpha4)以及干扰素诱导基因如Mx蛋白和ISG-15的表达与对照组无差异。rIHNV-Gvhsv GFP不像在接种IHNV的鱼细胞系中观察到的那样抑制细胞凋亡。这些研究表明,重组rIHNV-Gvhsv GFP作为一种活重组疫苗是一个有前景的候选者,并且还为进一步研究病毒致病性以及针对这些病毒感染的保护的分子基础提供了一个良好的模型。