O'Farrell Caroline, Vaghefi Nikta, Cantonnet Monique, Buteau Bénédicte, Boudinot Pierre, Benmansour Abdenour
Unité de Virologie et Immunologie Moléculaires, Institut National de la Recherche Agronomique, 78352 Jouy-en-Josas Cedex, France.
J Virol. 2002 Aug;76(16):8040-9. doi: 10.1128/jvi.76.16.8040-8049.2002.
Virus infections induce changes in the expression of host cell genes. A global knowledge of these modifications should help to better understand the virus/host cell interactions. To obtain a more comprehensive view of the rainbow trout response to a viral infection, we used the subtractive suppressive hybridization methodology in the viral hemorrhagic septicemia model of infection. We infected rainbow trout leukocytes with viral hemorrhagic septicemia virus (VHSV), and total RNA from infected and mock-infected cells was compared at 40 h postinfection. Twenty-four virus-induced genes were ultimately retrieved from the subtracted cDNA library, and their differential expression was further confirmed by semiquantitative reverse transcription-PCR and Northern blot analysis. Among these sequences, three were already described as VHSV-induced genes. Eight sequences with known homologs were extended to full-length cDNA using 5' and 3' rapid amplification of cDNA ends, and they were subsequently divided into three functional subsets. Four genes were homologous to mammalian interferon responsive genes, three were similar to chemo-attractant molecules (CXC chemokine, galectin), and two had nucleic acid binding domains. All of the virus-induced genes were also induced by rainbow trout interferon, indicating that the interferon pathway is the predominant component of the anti-VHSV response. They were also expressed in vivo in experimentally infected fish, indicating their biological relevance in natural infection.
病毒感染会诱导宿主细胞基因表达发生变化。全面了解这些修饰有助于更好地理解病毒与宿主细胞之间的相互作用。为了更全面地了解虹鳟对病毒感染的反应,我们在病毒性出血性败血症感染模型中采用了抑制性消减杂交方法。我们用病毒性出血性败血症病毒(VHSV)感染虹鳟白细胞,并在感染后40小时比较感染细胞和模拟感染细胞的总RNA。最终从消减cDNA文库中获得了24个病毒诱导基因,其差异表达通过半定量逆转录PCR和Northern印迹分析进一步得到证实。在这些序列中,有三个已被描述为VHSV诱导基因。利用5'和3' cDNA末端快速扩增技术将八个具有已知同源物的序列扩展为全长cDNA,随后将它们分为三个功能亚组。四个基因与哺乳动物干扰素反应基因同源,三个与趋化因子分子(CXC趋化因子、半乳糖凝集素)相似,两个具有核酸结合结构域。所有病毒诱导基因也被虹鳟干扰素诱导,这表明干扰素途径是抗VHSV反应的主要组成部分。它们也在实验感染的鱼体内表达,表明它们在自然感染中的生物学相关性。