Molecular characterization and immune functional analysis of IRF2 in common carp (Cyprinus carpio L.): different regulatory role in the IFN and NF-κB signalling pathway

Abstract Background Interferon regulatory factor 2 (IRF2) is an important transcription factor, which can regulate the IFN response and plays a role in antiviral innate immunity in teleost. Results In the present study, the full-length cDNA sequence of IRF2 (CcIRF2) was characterized in common carp (Cyprinus carpio L.), which encoded a protein containing a conserved DNA-binding domain (DBD) and an IRF-associated domain (IAD). Phylogenetic analysis showed that CcIRF2 was most closely related with IRF2 of Ctenopharyngodon idella. CcIRF2 transcripts were detectable in all examined tissues, with higher expression in the gills, spleen and brain. CcIRF2 expression was upregulated in immune-related tissues of common carp upon polyinosinic:polycytidylic acid (poly (I:C)) and Aeromonas hydrophila stimulation and induced by poly (I:C), lipopolysaccharide (LPS), peptidoglycan (PGN) and flagellin in the peripheral blood leucocytes (PBLs) and head kidney leukocytes (HKLs). In addition, overexpression of CcIRF2 decreased the expression of IFN and IFN-stimulated genes (ISGs), and a dual-luciferase reporter assay revealed that CcIRF2 could increase the activation of NF-κB. Conclusions These results indicate that CcIRF2 participates in antiviral and antibacterial immune response and negatively regulates the IFN response, which provide a new insight into the regulation of IFN system in common carp, and are helpful for the prevention and control of infectious diseases in carp farming.

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Identification of an IRF10 gene in common carp (Cyprinus carpio L.) and analysis of its function in the antiviral and antibacterial immune response

BMC Veterinary Research ◽

10.1186/s12917-020-02674-z ◽

2020 ◽

Vol 16 (1) ◽

Author(s):

Yaoyao Zhu ◽

Shijuan Shan ◽

Huaping Zhao ◽

Rongrong Liu ◽

Hui Wang ◽

...

Keyword(s):

Common Carp ◽

Cyprinus Carpio ◽

Head Kidney ◽

Ctenopharyngodon Idella ◽

Poly I:C ◽

Type I ◽

Regulatory Factors ◽

Polyinosinic:Polycytidylic Acid ◽

Transcriptional Regulatory ◽

Localization Signal

Abstract Background Interferon (IFN) regulatory factors (IRFs), as transcriptional regulatory factors, play important roles in regulating the expression of type I IFN and IFN- stimulated genes (ISGs) in innate immune responses. In addition, they participate in cell growth and development and regulate oncogenesis. Results In the present study, the cDNA sequence of IRF10 in common carp (Cyprinus carpio L.) was characterized (abbreviation, CcIRF10). The predicted protein sequence of CcIRF10 shared 52.7–89.2% identity with other teleost IRF10s and contained a DNA-binding domain (DBD), a nuclear localization signal (NLS) and an IRF-associated domain (IAD). Phylogenetic analysis showed that CcIRF10 had the closest relationship with IRF10 of Ctenopharyngodon idella. CcIRF10 transcripts were detectable in all examined tissues, with the highest expression in the gonad and the lowest expression in the head kidney. CcIRF10 expression was upregulated in the spleen, head kidney, foregut and hindgut upon polyinosinic:polycytidylic acid (poly I:C) and Aeromonas hydrophila stimulation and induced by poly I:C, lipopolysaccharide (LPS) and peptidoglycan (PGN) in peripheral blood leucocytes (PBLs) and head kidney leukocytes (HKLs) of C. carpio. In addition, overexpression of CcIRF10 was able to decrease the expression of the IFN and IFN-stimulated genes PKR and ISG15. Conclusions These results indicate that CcIRF10 participates in antiviral and antibacterial immunity and negatively regulates the IFN response, which provides new insights into the IFN system of C. carpio.

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Identification of an IRF10 gene in common carp (Cyprinus carpio L.) and analysis of its function in the antiviral and antibacterial immune response

10.21203/rs.3.rs-63536/v2 ◽

2020 ◽

Author(s):

Yaoyao Zhu ◽

Shijuan Shan ◽

Huaping Zhao ◽

Rongrong Liu ◽

Hui Wang ◽

...

Keyword(s):

Common Carp ◽

Cyprinus Carpio ◽

Head Kidney ◽

Ctenopharyngodon Idella ◽

Poly I:C ◽

Type I ◽

Regulatory Factors ◽

Common Carp Cyprinus Carpio ◽

Transcriptional Regulatory ◽

Carp Cyprinus Carpio

Abstract Background: Interferon (IFN) regulatory factors (IRFs), as transcriptional regulatory factors, play important roles in regulating the expression of type I IFN and IFN stimulated genes (ISGs) in innate immune responses. In addition, they participate in cell growth and development and regulate oncogenesis. Results: In the present study, the cDNA sequence of IRF10 in common carp (Cyprinus carpio L.) was characterized (abbreviation, CcIRF10). The predicted protein sequence of CcIRF10 shared 52.7-89.2% identity with other teleost IRF10s and contained a DNA-binding domain (DBD), a nuclear localization signal (NLS) and an IRF-associated domain (IAD). Phylogenetic analysis showed that CcIRF10 had the closest relationship with IRF10 of Ctenopharyngodon idella. CcIRF10 transcripts were detectable in all examined tissues, with the highest expression in the gonad and the lowest expression in the head kidney. CcIRF10 expression was upregulated in the spleen, head kidney, foregut and hindgut upon polyinosinic:polycytidylic acid (poly I:C) and Aeromonas hydrophila stimulation and induced by poly I:C, lipopolysaccharide (LPS) and peptidoglycan (PGN) in peripheral blood leucocytes (PBLs) and head kidney leukocytes (HKLs) of C. carpio. In addition, overexpression of CcIRF10 was able to decrease the expression of the IFN and IFN stimulated genes PKR and ISG15. Conclusions: These results indicate that CcIRF10 participates in antiviral and antibacterial immunity and negatively regulates the IFN response, which provides new insights into the IFN system of C. carpio.

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Identification and functional characterization of a fish-specific tlr19 in common carp (Cyprinus carpio L.) that recruits TRIF as an adaptor and induces ifn expression during the immune response

Veterinary Research ◽

10.1186/s13567-021-00957-3 ◽

2021 ◽

Vol 52 (1) ◽

Author(s):

Shijuan Shan ◽

Rongrong Liu ◽

Hanxiao Feng ◽

Fei Meng ◽

Muhanmmad Aizaz ◽

...

Keyword(s):

Common Carp ◽

Cyprinus Carpio ◽

Critical Role ◽

Functional Characterization ◽

Head Kidney ◽

Poly I:C ◽

Future Research ◽

Free Ribosome ◽

Early Endosomes ◽

Localization Analysis

AbstractToll-like receptor 19 (Tlr19) is a fish-specific TLR that plays a critical role in innate immunity. In the present study, we aimed to identify tlr19 from common carp (Cyprinus carpio L.) and explored its expression profile, localization, adaptor, and signaling pathways. A novel tlr19 cDNA sequence (Cctlr19) was identified in common carp. Phylogenetic analysis revealed that CcTlr19 was most closely related to Danio rerio Tlr19. Subcellular localization analysis indicates that CcTlr19 was synthesized in the free ribosome and then transported to early endosomes. Cctlr19 was constitutively expressed in all the examined tissues, with the highest expression in the brain. After poly(I:C) and Aeromonas hydrophila injection, the expression of Cctlr19 was significantly upregulated in immune-related organs. In addition, the expression of Cctlr19 was upregulated in head kidney leukocytes (HKL) upon stimulation with different ligands. Immunofluorescence and luciferase analyses indicate that CcTlr19 recruited TRIF as an adaptor. Furthermore, CcTlr19 can activate the expression of ifn-1 and viperin. Taken together, these findings lay the foundation for future research to investigate the mechanisms underlying fish tlr19.

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Identification and Functional Analysis of A Non-Mammalian IRF10 in Common Carp (Cyprinus Carpio. L) in The Antiviral and Antibacterial Immune Response

10.21203/rs.3.rs-63536/v1 ◽

2020 ◽

Author(s):

Yaoyao Zhu ◽

Shijuan Shan ◽

Huaping Zhao ◽

Rongrong Liu ◽

Hui Wang ◽

...

Keyword(s):

Common Carp ◽

Cyprinus Carpio ◽

Head Kidney ◽

Poly I:C ◽

Type I ◽

Innate Immune Responses ◽

Regulatory Factors ◽

Polycytidylic Acid ◽

Transcriptional Regulatory ◽

Localization Signal

Abstract Background: Interferon (IFN) regulatory factors (IRFs), as the transcriptional regulatory factors play important roles in regulating the expression of type I IFN and IFN-stimulated genes (ISGs) in innate immune responses, as well as participate in cell growth, development and regulating the oncogenesis.Results: In the present study, the cDNA sequence of IRF10 was characterized in common carp (Cyprinus carpio L.) (abbreviation, CcIRF10). The predicted protein sequence of CcIRF10 shared 52.7-89.2% identities to teleost IRF10 and contained a DNA-binding domain (DBD), a nuclear localization signal (NLS) and an IRF associated domain (IAD). Phylogenetic analysis showed that CcIRF10 had a closer relationship with the IRF10 of Ctenopharyngodon idella. CcIRF10 transcripts was detectable in all examined tissues, with the highest expression in the gonad and the lowest expression level in the head kidney. The CcIRF10 expression was up-regulated in the spleen, head kidney, foregut and hindgut upon polyinosinic: polycytidylic acid (poly I:C) and Aeromonas hydrophila stimulation, and also induced by poly I:C, LPS and PGN in the peripheral blood leucocytes (PBLs) and head kidney leukocytes (HKLs) of C. carpio. In addition, overexpression of CcIRF10 was able to decrease the expressions of IFN-related genes, PKR and ISG5.Conclusions: These results indicated that CcIRF10 participated in the antiviral and antibacterial immunity, and negatively regulated the IFN response, which provided new insights into the IFN system of C. carpio.

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Cyprinus carpio TRIF Participates in the Innate Immune Response by Inducing NF-κB and IFN Activation and Promoting Apoptosis

Frontiers in Immunology ◽

10.3389/fimmu.2021.725150 ◽

2021 ◽

Vol 12 ◽

Author(s):

Rongrong Liu ◽

Xiaoye Liu ◽

Meijiao Song ◽

Yue Qi ◽

Hua Li ◽

...

Keyword(s):

Immune Response ◽

Common Carp ◽

Innate Immune Response ◽

Cyprinus Carpio ◽

Critical Role ◽

Innate Immune ◽

Poly I:C ◽

Luciferase Reporter

TRIF, an important adaptor downstream of Toll-like receptor signaling, plays a critical role in the innate immune response. In this study, the full-length coding sequence of TRIF from common carp (Cyprinus carpio L.) was cloned and characterized. Bioinformatics analysis showed that common carp TRIF exhibited a conserved TIR domain and had the closest relationship with grass carp TRIF. Expression analysis revealed that TRIF was constitutively expressed in the examined tissues of common carp, with the highest expression in the spleen and the lowest expression in the head kidney, and could be upregulated under Aeromonas hydrophila and poly(I:C) stimulation in vivo and under poly(I:C), LPS, PGN, flagellin, and Pam3CSK4 stimulation in vitro. Laser confocal microscopy showed that common carp TRIF colocalized with the Golgi apparatus. A luciferase reporter assay showed that carp TRIF elicited the activity of ifn-1 and nf-κb through the C-terminal domain. Additionally, crystal violet staining and qPCR assays revealed that carp TRIF inhibited the replication of SVCV in epithelioma papulosum cyprini (EPC) cells. Then, the signaling downstream of carp TRIF was investigated. Coimmunoprecipitation and Western blotting analysis demonstrated that carp TRIF interacted with TBK1 and augmented the expression of TRAF6 and phosphorylation of TBK1. Overexpression of carp TRIF significantly enhanced the expression of interferon-stimulated genes and inflammatory cytokines. Furthermore, flow cytometric (FCM) analysis suggested that carp TRIF induced apoptosis through the activation of caspase-8. In summary, our study indicated that TRIF plays an essential role in the innate immune responses of common carp against bacterial and viral infection.

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Enrichment of common carp (Cyprinus carpio) diet with Malic acid: Effects on skin mucosal immunity, antioxidant defecne and growth performance

Annals of Animal Science ◽

10.2478/aoas-2020-0092 ◽

2020 ◽

Vol 0 (0) ◽

Author(s):

Roghieh Safari ◽

Seyed Hossein Hoseinifar ◽

Maryam Dadar ◽

Hien Van Doan

Keyword(s):

Common Carp ◽

Cyprinus Carpio ◽

Malic Acid ◽

Control Group ◽

Skin Mucus ◽

Antioxidant Genes ◽

Treatment Comparison ◽

Genes Expression ◽

Significant Difference ◽

And Control

AbstractThe present study investigated possible effects of dietary malic acid on the expression of immunity, antioxidant and growth related genes expression as well as skin mucus immune parameters in common carp. Common carp (Cyprinus carpio) fingerlings were fed diets supplemented with different levels (0 [control], 0.5%, 1%, 2%) of malic acid (MA) for 60 days. The results revealed highest expression levels of immune-related genes (tnf-alpha, il1b, il8 and lyz) in skin of common carp fed 2% MA (P < 0.05). Regarding 1% MA treatment comparison with control group, significant difference was noticed just in case of lyz (P < 0.05). Evaluation of growth related genes expression revealed no significant difference between treatments (P > 0.05). The study of antioxidant related genes (gsta and gpx) in common carp skin fed with MA, showed significant difference between treated groups and control (P < 0.05). Carps fed with 2% MA had highest alkaline phosphatase activity in skin mucus compared other treated groups and control (P < 0.05). There were no significant difference between 0.5% and 1% and control (P > 0.05). The study of total protein and total immunoglobulin (Ig) in common carp skin musus revealed no alteration following MA treatment (P > 0.05). The present data demonstrated that feeding with MA altered immune and antioxidant genes expression in skin mucus of common carp.

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Acetylcholinesterase enzyme activity in carp brain and muscle after acute exposure to diafuran

Scientia Agricola ◽

10.1590/s0103-90162008000400003 ◽

2008 ◽

Vol 65 (4) ◽

pp. 340-345 ◽

Cited By ~ 19

Author(s):

Jaqueline Ineu Golombieski ◽

Enio Marchesan ◽

Edinalvo Rabaioli Camargo ◽

Joseânia Salbego ◽

Joele Schmitt Baumart ◽

...

Keyword(s):

Common Carp ◽

Cyprinus Carpio ◽

Grass Carp ◽

Ache Activity ◽

Bighead Carp ◽

Ctenopharyngodon Idella ◽

Control Treatment ◽

Aristichthys Nobilis ◽

Early Biomarker ◽

The Common

Sublethal adverse effects may result from exposure of aquatic organisms to insecticides at environmentally relevant concentrations. Fingerlings of the common carp (Cyprinus carpio, Linnaeus, 1758), grass carp (Ctenopharyngodon idella, Valenciennes, 1844), and bighead carp (Aristichthys nobilis, Richardson, 1845) were exposed to diafuran, an insecticide widely used during rice cultivation in Southern Brazil. The aim of this study was to verify the relationship between the lethal concentration (LC50) of diafuran and the acetylcholinesterase (AChE) activity in brain and muscle tissues of these species as a possible early biomarker of exposure to this insecticide. LC50 was determined for fish exposed to diafuran concentrations during 96 h (short term): common carp: control, 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0 mg L-1; grass carp: control, 1.0, 2.0, 3.0 and 3.5 mg L-1 and, bighead carp: control, 0.5, 1.0, 1.5, 2.0, 3.0 and 4.0 mg L-1, as well as the determination of AChE at concentrations near LC50 for these species. LC50 values (nominal concentrations) were 1.81 mg L-1 for the common carp, 2.71 mg L-1 for the grass carp and, 2.37 mg L-1 for the bighead carp. All carps exposed to diafuran were lethargic (lower concentrations) or immobile. Diafuran inhibited the acetylcholinesterase activity in brain (~38%) and muscle (~50%) of all species. Muscle of bighead carp under control treatment showed higher specific AChE activity than brain (14.44 against 5.94 µmol min-1 g protein-1, respectively). Concentrations of diafuran used for rice cropping may affect Cyprinus carpio, Ctenopharyngodon idella and Aristichthys nobilis behaviors and the AChE activities in brain and muscle of these species may be an early biomarker of toxicity of this insecticide.

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