T cell transcripts and T cell activities in the gills of the teleost fish sea bass (Dicentrarchus labrax)

Nuñez-Ortiz, Noelia; Gerdol, Marco; Stocchi, V.; Marozzi, C.; Randelli, Elisa; Bernini, C.; Buonocore, Francesco; Picchietti, Simona; Papeschi, Cristiano; Sood, Neeraj; Pallavicini, Alberto; Scapigliati, Giuseppe

doi:10.1016/j.dci.2014.07.015

Cited by 59 publications

(28 citation statements)

References 45 publications

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“…Two different Mx genes had been identified in European sea bass, proving that the isoform MxA is particularly modulated in nodavirus-infected sea bass [20, 46]. As previously shown [47], the ISG12 gene can also be employed in sea bass as a marker of antiviral activities, since this ISG isoform is well expressed in gills [22], and significant ISG12 gene modulation can be induced in sea bass after in vitro stimulation with virus-mimicking substance poly I:C [47]. …”

Section: Discussionmentioning

confidence: 94%

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A formalin-inactivated immunogen against viral encephalopathy and retinopathy (VER) disease in European sea bass (Dicentrarchus labrax): immunological and protection effects

Nuñez-Ortiz

Pascoli

Picchietti

et al. 2016

Vet Res

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The European sea bass (Dicentrarchus labrax) is an important farmed fish species in the Mediterranean area, very sensitive to the infection by encephalopathy and retinopathy virus (VERv), or Betanodavirus, which causes massive mortalities. Effective vaccines to fight the pathology are not yet available and in this work we describe a promising intraperitoneal immunization route against VERv of sea bass juveniles. We performed intraperitoneal and immersion immunization trials with a VERv (isolate 283.2009 RGNNV) inactivated by formalin, β-propiolactone and heat treatment. Interestingly, the intraperitoneal immunization with formalin-inactivated VERv induced a significant antigen-specific IgM production, differently from other inactivation protocols. However, the same formalin-inactivated antigen resulted in very low IgM antibodies when administered by immersion. Following the intraperitoneal injection with formalin-inactivated virus, the quantitative expression of the antiviral MxA gene showed a modulation of transcripts in the gut after 48 h and on head kidney after 24 h, whereas ISG12 gene was significantly up-regulated after 48 h on both tissues. In immersion immunization with formalin-inactivated VERv, a modulation of MxA and ISG12 genes after 24 h post-treatment was detected in the gills. An effective uptake of VERv particles in the gills was confirmed by immunohistochemistry using anti-VERv antibodies. Lastly, in challenge experiments using live VERv after intraperitoneal immunization with formalin-inactivated VERv, we observed a significant increase (81.9%) in relative survival percentage with respect to non-immunized fish, whereas immersion immunization resulted in no protection. Our results suggest that intraperitoneal immunization with formalin-inactivated VERv could be a safe and effective strategy to fight Betanodavirus infection in European sea bass.Electronic supplementary materialThe online version of this article (doi:10.1186/s13567-016-0376-3) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 94%

“…Gills were fixed in ice-cold Bouin’s fixative for 7 h in Bouin’s at 4 °C. After embedding in paraplast, blocks were serially sectioned at a thickness of 7 μm as previously described [22]. The immunohistochemical reaction was performed by ABC-peroxidase with nickel enhancement, as previously described [23].…”

Section: Methodsmentioning

confidence: 99%

A formalin-inactivated immunogen against viral encephalopathy and retinopathy (VER) disease in European sea bass (Dicentrarchus labrax): immunological and protection effects

Nuñez-Ortiz

Pascoli

Picchietti

et al. 2016

Vet Res

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“…A partial secretory IgT nucleotide sequence was identified after the analysis of a sea bass gills transcriptome [31]. From this sequence two primers were designed (IGTFW 5′-GACCAACTGTGTTTCCTCTGATGC-3′ and IGTRV 5′-GCCATCCTTATAGATGCCAGGGGA-3′) and used in RT-PCR on total RNA extracted with Trisure (Bio-line) solution from gills of a juvenile sea bass (200 g of weight).…”

Section: Methodsmentioning

confidence: 99%

Immunoglobulin T from sea bass (Dicentrarchus labrax L.): molecular characterization, tissue localization and expression after nodavirus infection

et al. 2017

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BackgroundImmunoglobulins (Igs) are fundamental components of the adaptive immune system of vertebrates, with the IgT/IgZ isotype specific of Teleosts. In this paper we describe the identification of an IgT heavy chain from the European sea bass (Dicentrarchus labrax L.), its molecular characterization and tissue mRNA localization by in situ hybridization.ResultsSea bass IgT consists of 552 aa (Accession Number KM410929) and it contains a putative 19 amino acids long signal peptide and one potential N-glycosylation site. The C-region consists of four CH domains; each contains the cysteine and tryptophan residues required for their correct folding. Based on the recent sequencing of sea bass genome, we have identified five different genomic contigs bearing exons unequivocally pertaining to IgT (CH2, CH3 and CH4), but none corresponded to a complete IgH locus as IgT sequences were found in the highly fragmented assembled genomic regions which could not be assigned to any major scaffold. The 3D structure of sea bass IgT has been modelled using the crystal structure of a mouse Ig gamma as a template, thus showing that the amino acid sequence is suitable for the expected topology referred to an immunoglobulin-like architecture. The basal expression of sea bass IgT and IgM in different organs has been analysed: gut and gills, important mucosal organs, showed high IgT transcripts levels and this was the first indication of the possible involvement of sea bass IgT in mucosal immune responses. Moreover, sea bass IgT expression increased in gills and spleen after infection with nodavirus, highlighting the importance of IgT in sea bass immune responses. In situ hybridization confirmed the presence of IgT transcripts in the gut and it revealed a differential expression along the intestinal tract, with a major expression in the posterior intestine, suggesting the hindgut as a site for the recruitment of IgT+ cells in this species. IgT transcripts were also found in gill filaments and parallel lamellae and, for the first time, we identified scattered IgT positive cells in the liver, with a strong signal in the hepatic parenchyma.ConclusionsIn conclusion, we performed a full molecular characterization of IgT in sea bass that points out its possible involvement in mucosal immune responses of this species.

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“…In European sea bass, massive gene expression analysis have undertaken for gills, liver, brain, eggs, and larvae (Geay et al, 2011; Magnanou et al, 2014; Nuñez Ortiz et al, 2014; Kaitetzidou et al, 2015). A high-quality chromosome-scale genome assembly is also available (Tine et al, 2014) for this species, but in-depth analysis of the intestinal transcriptome remains to be fully addressed.…”

Section: Introductionmentioning

confidence: 99%

Gene Expression Profiling Reveals Functional Specialization along the Intestinal Tract of a Carnivorous Teleostean Fish (Dicentrarchus labrax)

2016

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High-quality sequencing reads from the intestine of European sea bass were assembled, annotated by similarity against protein reference databases and combined with nucleotide sequences from public and private databases. After redundancy filtering, 24,906 non-redundant annotated sequences encoding 15,367 different gene descriptions were obtained. These annotated sequences were used to design a custom, high-density oligo-microarray (8 × 15 K) for the transcriptomic profiling of anterior (AI), middle (MI), and posterior (PI) intestinal segments. Similar molecular signatures were found for AI and MI segments, which were combined in a single group (AI-MI) whereas the PI outstood separately, with more than 1900 differentially expressed genes with a fold-change cutoff of 2. Functional analysis revealed that molecular and cellular functions related to feed digestion and nutrient absorption and transport were over-represented in AI-MI segments. By contrast, the initiation and establishment of immune defense mechanisms became especially relevant in PI, although the microarray expression profiling validated by qPCR indicated that these functional changes are gradual from anterior to posterior intestinal segments. This functional divergence occurred in association with spatial transcriptional changes in nutrient transporters and the mucosal chemosensing system via G protein-coupled receptors. These findings contribute to identify key indicators of gut functions and to compare different fish feeding strategies and immune defense mechanisms acquired along the evolution of teleosts.

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T cell transcripts and T cell activities in the gills of the teleost fish sea bass (Dicentrarchus labrax)

Cited by 59 publications

References 45 publications

A formalin-inactivated immunogen against viral encephalopathy and retinopathy (VER) disease in European sea bass (Dicentrarchus labrax): immunological and protection effects

A formalin-inactivated immunogen against viral encephalopathy and retinopathy (VER) disease in European sea bass (Dicentrarchus labrax): immunological and protection effects

Immunoglobulin T from sea bass (Dicentrarchus labrax L.): molecular characterization, tissue localization and expression after nodavirus infection

Gene Expression Profiling Reveals Functional Specialization along the Intestinal Tract of a Carnivorous Teleostean Fish (Dicentrarchus labrax)

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