Identification and expression analysis of two interleukin-23α (p19) isoforms, in rainbow trout Oncorhynchus mykiss and Atlantic salmon Salmo salar

Developmental & Comparative Immunology

Scott

Secombes

2016

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Flagellin is the principal component of bacterial flagellum and a major target of the host immune system. To provide new insights into the role of flagellin in fish immune responses to flagellated microorganisms, a recombinant flagellin from Yersinia ruckeri (rYRF) was produced and its bioactivity investigated in the trout macrophage cell line RTS-11 and head kidney cells. rYRF is a potent activator of pro-inflammatory cytokines, acute phase proteins, antimicrobial peptides and subunits of the IL-12 cytokine family. This and the synergy seen with IFN-γ to enhance further expression of specific IL-12 and TNF-α isoforms may suggest that flagellin could be a useful immune stimulant or adjuvant for use in aquaculture. Gene paralogues were often differentially modulated, highlighting the need to study all of the paralogues of immune genes in fish to gain a full understanding of the effects of PAMPs or other stimulants, and the potential immune responses elicited.

Section: S2b-d)mentioning

confidence: 99%

Re-examination of the rainbow trout (Oncorhynchus mykiss) immune response to flagellin: Yersinia ruckeri flagellin is a potent activator of acute phase proteins, anti-microbial peptides and pro-inflammatory cytokines in vitro

Developmental & Comparative Immunology

Scott

Secombes

2016

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“…With the recent release of salmonid genomes [19][20] and the identification of a large number of cytokine genes including the paralogues for IL-1β [21], TNF-α [22], IL-17A/F [23][24], IL-4/13 [25] and subunits of the IL-12 family [26][27][28][29] in salmonids, it is timely to revisit the early immune responses to ERM vaccination.…”

Section: Introductionmentioning

confidence: 99%

Dissecting the immune pathways stimulated following injection vaccination of rainbow trout (Oncorhynchus mykiss) against enteric redmouth disease (ERM)

Secombes

Fish & Shellfish Immunology

2019

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Enteric redmouth disease (ERM or yersiniosis) is one of the most important diseases of salmonids and leads to significant economic losses. It is caused by the Gram-negative bacterium Yersinia ruckeri but can be controlled by bacterin vaccination. The first commercial ERM vaccine was licenced in 1976 and is one of the most significant and successful health practices within the aquaculture industry. Although ERM vaccination provides complete protection, knowledge of the host immune response to the vaccine and the molecular mechanisms that underpin the protection elicited is limited. In this report, we analysed the expression in spleen and gills of a large set of genes encoding for cytokines, acute phase proteins (APPs) and antimicrobial peptides (AMPs) in response to ERM vaccination in rainbow trout, Oncorhynchus mykiss. Many immune genes in teleost fish are known to have multiple paralogues that can show differential responses to ERM vaccination, highlighting the necessity to determine whether all of the genes present react in a similar manner. ERM vaccination immediately activated a balanced inflammatory response with correlated expression of both pro- and anti-inflammatory cytokines (eg IL-1β1-2, TNF-α1-3, IL-6, IL-8 and IL-10A etc.) in the spleen. The increase of pro-inflammatory cytokines may explain the systemic upregulation of APPs (eg serum amyloid A protein and serum amyloid protein P) and AMPs (eg cathelicidins and hepcidin) seen in both spleen and gills. We also observed an upregulation of all the α-chains but only one β-chain (p40B2) of the IL-12 family cytokines, that suggests specific IL-12 and IL-23 isoforms with distinct functions might be produced in the spleen of vaccinated fish. Notably the expression of Th1 cytokines (IFN-γ1-2) and a Th17 cytokine (IL-17A/F1a) was also up-regulated and correlated with enhanced expression of the IL-12 family α-chains, and the majority of pro- and anti-inflammatory cytokines, APPs and AMPs. These expression profiles may suggest that ERM vaccination activates host innate immunity and expression of specific IL-12 and IL-23 isoforms leading to a Th1 and Th17 biased immune response. A late induction of Th2 cytokines (IL-4/13B1-2) was also observed, that may have a homeostatic role and/or involvement in antibody production. This study has increased our understanding of the host immune response to ERM vaccination and the adaptive pathways involved. The early responses of a set of genes established in this study may provide essential information and function as biomarkers in future vaccine development in aquaculture.

“…Primers (Table 1) were designed to the 5'and 3'untranslated regions (UTR) and used for PCR amplification and cloning of the complete coding region using a mixed tissue cDNA sample. The cloning, DNA and protein sequence analysis was as described previously [19,20]. Briefly, the nucleotide sequences generated were assembled and analysed using the AlignIR™ Software (LI-COR, Inc.).…”

Section: Cloning Of Arginase Isoforms and Sequence Analysismentioning

confidence: 99%

“…Six rainbow trout (average weight, 142 g) were anaesthetised using 2-phenoxyethanol (Sigma-Aldrich, UK), killed and 17 tissues (tail fin, adipose fin, gills, thymus, brain, scales, skin, muscle, adipose tissue, liver, spleen, gonad, head kidney, posterior kidney, intestine, heart and blood) were collected. RNA extraction and complementary (c) DNA synthesis was as described previously [19,20].…”

Section: Constitutive Expression Of Rainbow Trout Arginase Isoforms Imentioning

confidence: 99%

Characterisation of arginase paralogues in salmonids and their modulation by immune stimulation/ infection

Benedicenti

Fish & Shellfish Immunology

et al. 2017

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In this study we show that four arginase isoforms (arg1a, arg1b, arg2a, arg2b) exist in rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar). We have characterised these molecules in terms of a) sequence analysis, b) constitutive expression in different tissues, and modulated expression following c) stimulation of head kidney macrophages in vitro, or d) vaccination/infection with Yersinia ruckeri and e) parasite infection (AGD caused by Paramoeba perurans and PKD caused by Tetracapsuloides bryosalmonae). Synteny analysis suggested that these arginase genes are paralogues likely from the Ss4R duplication event, and amino acid identity/similarity analyses showed that the proteins are relatively well conserved across species. In rainbow trout constitutive expression of one or both paralogues was seen in most tissues but different constitutive expression patterns were observed for the different isoforms. Stimulation of rainbow trout head kidney macrophages with PAMPs and cytokines also revealed isoform specific responses and kinetics, with arg1a being particularly highly modulated by the PAMPs and pro-inflammatory cytokines. In contrast the type II arginase paralogues were induced by rIl-4/13, albeit to a lesser degree. Vaccination and infection with Y. ruckeri also revealed isoform specific responses, with variation in tissue expression level and kinetics. Lastly, the impact of parasite infection was studied, where down regulation of arg1a and arg1b was seen in two different models (AGD in salmon and PKD in trout) and of arg2a in AGD. The differential responses seen are discussed in the context of markers of type II responses in fish and paralogue subfunctionalization.