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*Instituto de Investigaciones Marinas, Consejo Superior de Investigaciones Científicas (CSIC), Vigo, Spain;
Center of Marine Biotechnology, University of Maryland Biotechnology Institute, Baltimore, MD 21202; and
Instituto de Biología Molecular y Celular, Universidad Miguel Hernández, Elche (Alicante), Spain
Sea bass nervous necrosis virus is the causative agent of viral nervous necrosis, a disease responsible of high economic losses in larval and juvenile stages of cultured sea bass (Dicentrarchus labrax). To identify genes potentially involved in antiviral immune defense, gene expression profiles in response to nodavirus infection were investigated in sea bass head kidney using the suppression subtractive hybridization (SSH) technique. A total of 8.7% of the expressed sequence tags found in the SSH library showed significant similarities with immune genes, of which a prototype galectin (Sbgalectin-1), two C-type lectins (SbCLA and SbCLB) from groups II and VII, respectively, and a short pentraxin (Sbpentraxin) were selected for further characterization. Results of SSH were validated by in vivo up-regulation of expression of Sbgalectin-1, SbCLA, and SbCLB in response to nodavirus infection. To examine the potential role(s) of Sbgalectin-1 in response to nodavirus infection in further detail, the recombinant protein (rSbgalectin-1) was produced, and selected functional assays were conducted. A dose-dependent decrease of respiratory burst was observed in sea bass head kidney leukocytes after incubation with increasing concentrations of rSbgalectin-1. A decrease in IL-1β, TNF-
, and Mx expression was observed in the brain of sea bass simultaneously injected with nodavirus and rSbgalectin-1 compared with those infected with nodavirus alone. Moreover, the protein was detected in the brain from infected fish, which is the main target of the virus. These results suggest a potential anti-inflammatory, protective role of Sbgalectin-1 during viral infection.
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
1 This research was supported by the European Union project (SSP8-CT-2003-501984), the project CSD2007-00002 Aquagenomics funded by the program Consolider-Ingenio 2010 from the Spanish Ministerio de Educación y Ciencia and Xunta de Galicia (PGIDIT04PXIC40203PM). L.P.-B. and J.A.-G. were supported by Formación de personal universitario fellowships from Ministerio de Educación y Ciencia (Spain). Work at the Center of Marine Biotechnology, University of Maryland Biotechnology Institute, was supported by grants R01 GM070589-01 from the National Institutes of Health, and IOS-0822257 from the National Science Foundation (to G.R.V.).
2 The sequences presented in this article have been submitted to GenBank (www.ncbi. nlm.nih.gov/Genbank) under accession numbers as follows: Genomic Sbgalectin-1, EU660934; Sbgalectin-1, EU660937; SbCLA, EU660935; SbCLB, EU660936; Sbpentraxin, EU660933; and ESTs, from FF578829 to FF579034.
3 Address correspondence and reprint requests to Dr. Beatriz Novoa, Instituto de Investigaciones Marinas, Consejo Superior de Investigaciones Científicas (CSIC), Eduardo Cabello, 6. 36208 Vigo, Spain. E-mail address: virus{at}iim.csic.es
4 Abbreviations used in this paper: SBNNV, Sea bass nervous necrosis virus; CRD, carbohydrate-recognition domain; CTLD, C-type-like domain; EST, expressed sequence tag; ORF, open reading frame; RLU, relative luminescence unit; SSH, suppression subtractive hybridization; qPCR, quantitative PCR.
5 The online version of this article contains supplemental material.
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