The DSpace digital repository system captures, stores, indexes, preserves, and distributes digital research material. http://http://dspace.unitus.it:80 2013-05-19T23:32:24Z 2013-05-19T23:32:24Z Scapigliati, Giuseppe Buonocore, Francesco Randelli, Elisa Casani, Daniela Meloni, Sabrina Zarletti, Gianpaolo Tiberi, M. Pietretti, D. Boschi, I. Manchado, Manuel Martin-Antonio, B. Jimenez-Cantizano, R. Bovo, Giuseppe Borghesan, Fabio Lorenzen, Niels Einer-Jensen, Katja Adams, S. Thompson, K. Alonso, C. Béjar, Julia Cano, Irene Borrego, Juan J. Alvarez, M.Carmen http://hdl.handle.net/2067/1685 2011-06-28T10:07:01Z 2010-12-31T23:00:00Z

Title: Cellular and molecular immune responses of the sea bass (Dicentrarchus labrax) experimentally infected with betanodavirus Authors: Scapigliati, Giuseppe; Buonocore, Francesco; Randelli, Elisa; Casani, Daniela; Meloni, Sabrina; Zarletti, Gianpaolo; Tiberi, M.; Pietretti, D.; Boschi, I.; Manchado, Manuel; Martin-Antonio, B.; Jimenez-Cantizano, R.; Bovo, Giuseppe; Borghesan, Fabio; Lorenzen, Niels; Einer-Jensen, Katja; Adams, S.; Thompson, K.; Alonso, C.; Béjar, Julia; Cano, Irene; Borrego, Juan J.; Alvarez, M.Carmen Abstract: Naïve sea bass juveniles (38.4 ± 4.5 g) were intraperitoneally infected with a sublethal dose of betanodavirus isolate 378/I03, followed after 43 days by a similar boosting. This infection resulted in an overall mortality of 7.6 %. At various intervals, sampling of fish tissues were performed to investigate: i) B and T lymphocyte content in organs and tissues; ii), proliferation of leucocytes re-stimulated in vitro with inactivated virus; iii) presence of serum antibody specific for betanodavirus; iv) expression of genes coding for the following immunoregulatory molecules involved in innate and acquired responses: IFN-1, Mx, IL-1, Cox-2; IL-10, TGF-, TCR, CD4, CD8, IgM, by using a quantitative PCR array system developed for sea bass. The obtained results showed a detectable increase of T cells and B cells in PBL during betanodavirus infection. Furthermore, leucocytes obtained from blood, head kidney, and gills showed a detectable in “vitro” increase in viability upon addition of inactivated viral particles, as determined by measuring intracellular ATP concentration. ELISA analysis of sera showed that exposure to nodavirus induced a low, but specific antibody titer measured 43 days after infection, despite the presence of measurable levels of natural antibody. Finally, a strong upregulation of genes coding for IFN-1, Mx, and IgM was identified after both infection and boosting. Interestingly, an upregulation of Cox-2 until boosting, and of TGF- and IL-10 after boosting was also observed, while the other tested genes did not show any significant variations with respect to mock-treated fish. Overall, our work represents a first comprehensive analysis of cellular and molecular immune parameters in a fish species exposed to a pathogenic virus. Description: L'articolo è disponibile sul sito dell'editore http://www.sciencedirect.com/

2010-12-31T23:00:00Z Buonocore, Francesco Randelli, Elisa Lorenzen, Niels Einer-Jensen, Katja Scapigliati, Giuseppe http://hdl.handle.net/2067/2180 2011-08-03T11:05:42Z 2010-12-31T23:00:00Z

Title: Analysis of the expression and modulation of selected immune-related gene transcripts in the DLEC cell line from European sea bass (Dicentrarchus labrax) Authors: Buonocore, Francesco; Randelli, Elisa; Lorenzen, Niels; Einer-Jensen, Katja; Scapigliati, Giuseppe Abstract: Cell lines have been established from different fish species especially for virus isolation and for studying cell-pathogen interactions, and therefore are of interest in aquaculture. In this paper, we have investigated the presence and the regulation of some immune genes in the DLEC (Dicentrarchus labrax embryonic cells) cell line from European sea bass (Dicentrarchus labrax L.) to preliminary elucidate their action. The basal expression of the selected genes (interleukin- 1β (IL-1β), cyclooxygenase-2 (COX-2), transforming growth factor-β (TGF-β), CD8-α, major histocompatibility complex II-β (MHC II-β), interferon (IFN) and Mx protein (Mx)) have been investigated and, successively, their modulation have been studied both after stimulation with different mitogen agents and after a transfection with a sequence codifying for the coat protein of a fish nervous necrosis virus (NNV). The results have evidenced that the inflammatory molecules (IL-1β, COX-2, TGF-β), constitutively expressed by the DLEC cell line, are not up-regulated by the stimulation with lipopolysaccharide (LPS) from E. coli, whether the expression of the T-cell marker transcripts (CD8-α, MHC II-β) is influenced by the action of a lectin from Phaseolus vulgaris (PHA-L). Finally, the expression of the coat NNV protein in the DLEC cell line, after the transfection, led to an high up-regulation of IFN and Mx gene transcripts. These data suggest that the DLEC cell line recognize specific pathogen-associated molecular patterns (PAMPs) and, therefore, could be useful for studying T-cell pathways and viral responses in sea bass avoiding the use of live test animals.

2010-12-31T23:00:00Z