Structural and Functional Analyses of Type I IFNa Shed Light Into Its Interaction With Multiple Receptors in Fish

Wang, Zixuan; Xu, Jing; Feng, Jianhua; Wu, Kaizheng; Chen, Kangyong; Zhao, Jia; Zhu, Xiao-Zhen; Huang, Wei‐Ping; Liu, Qin; Wang, Bangjie; Chen, Xinhua; Wang, Junya; Zou, Jun

doi:10.3389/fimmu.2022.862764

Cited by 7 publications

(5 citation statements)

References 47 publications

(69 reference statements)

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“…The use of poly(I:C)to study antiviral immune mechanisms is commonly used, and the studies have shown that genes linked to viral immunity are upregulated when poly(I:C)is exposed with fish, fish leukocytes or fish cell lines ( 16 , 27 , 28 ). According to functional studies and the discovery of fish genes involved in the antiviral response, the IFN antiviral system is diverse among vertebrates ( 29 – 31 ). By producing cytokines, PRRs that recognize PAMPs ensure that the immune response elicited is specific to the invading pathogen, in contrast to e.g.…”

Section: Introductionmentioning

confidence: 99%

Transcriptome-wide analyses of early immune responses in lumpfish leukocytes upon stimulation with poly(I:C)

Rao,

Lunde,

Dolan

et al. 2023

Front. Immunol.

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BackgroundBoth bacterial and viral diseases are a major threat to farmed fish. As the antiviral immune mechanisms in lumpfish (Cyclopterus lumpus L.) are poorly understood, lumpfish leukocytes were stimulated with poly(I:C), a synthetic analog of double stranded RNA, which mimic viral infections, and RNA sequencing was performed.MethodsTo address this gap, we stimulated lumpfish leukocytes with poly(I:C) for 6 and 24 hours and did RNA sequencing with three parallels per timepoint. Genome guided mapping was performed to define differentially expressed genes (DEGs).ResultsImmune genes were identified, and transcriptome-wide analyses of early immune responses showed that 376 and 2372 transcripts were significantly differentially expressed 6 and 24 hours post exposure (hpe) to poly(I:C), respectively. The most enriched GO terms when time had been accounted for, were immune system processes (GO:0002376) and immune response (GO:0006955). Analysis of DEGs showed that among the most highly upregulated genes were TLRs and genes belonging to the RIG-I signaling pathway, including LGP2, STING and MX, as well as IRF3 and IL12A. RIG-I was not identified, but in silico analyses showed that genes encoding proteins involved in pathogen recognition, cell signaling, and cytokines of the TLR and RIG-I signaling pathway are mostly conserved in lumpfish when compared to mammals and other teleost species.ConclusionsOur analyses unravel the innate immune pathways playing a major role in antiviral defense in lumpfish. The information gathered can be used in comparative studies and lay the groundwork for future functional analyses of immune and pathogenicity mechanisms. Such knowledge is also necessary for the development of immunoprophylactic measures for lumpfish, which is extensively cultivated for use as cleaner fish in the aquaculture for removal of sea lice from Atlantic salmon (Salmo salar L.).

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Section: Introductionmentioning

confidence: 99%

Transcriptome-wide analyses of early immune responses in lumpfish leukocytes upon stimulation with poly(I:C)

Rao,

Lunde,

Dolan

et al. 2023

Front. Immunol.

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“…Following the removal of 50 ml for Western blot analysis, 40 ml of a-Myc-conjugated magnetic beads (Abmart) was added to the remaining supernatant and incubated overnight on a shaker. The magnetic beads were collected by centrifugation at 1,500 g for 3 min at 4°C, washed 3 times with ice-cold PBS, resuspended in 30 µl 2 × SDS loading buffer and prepared for Western blot analysis as detailed description previous (26). Briefly, Protein samples are separated by SDS-PAGE, then transferred to PVDF membranes using a semi-dry method.…”

Section: Co-immunoprecipitationmentioning

confidence: 99%

The first crystal structure of CD8αα from a cartilaginous fish

et al. 2023

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IntroductionCartilaginous fishes are the most evolutionary-distant vertebrates from mammals and possess an immunoglobulin (Ig)- and T cell-mediated adaptive immunity. CD8 is the hallmark receptor of cytotoxic T cells and is required for the formation of T cell receptor-major histocompatibility complex (TCR-MHC) class I complexes.MethodsRACE PCR was used to obtain gene sequences. Direct dilution was applied for the refolding of denatured recombinant CD8 protein. Hanging-drop vapor diffusion method was performed for protein crystallization.ResultsIn this study, CD8α and CD8β orthologues (termed ScCD8α and ScCD8β) were identified in small-spotted catshark (Scyliorhinus canicula). Both ScCD8α and ScCD8β possess an extracellular immunoglobulin superfamily (IgSF) V domain as in previously identified CD8 proteins. The genes encoding CD8α and CD8β are tandemly linked in the genomes of all jawed vertebrates studied, suggesting that they were duplicated from a common ancestral gene before the divergence of cartilaginous fishes and other vertebrates. We determined the crystal structure of the ScCD8α ectodomain homodimer at a resolution of 1.35 Å and show that it exhibits the typical topological structure of CD8α from endotherms. As in mammals, the homodimer formation of ScCD8αα relies upon interactions within a hydrophobic core although this differs in position and amino acid composition. Importantly, ScCD8αα shares the canonical cavity required for interaction with peptide-loaded MHC I in mammals. Furthermore, it was found that ScCD8α can co-immunoprecipitate with ScCD8β, indicating that it can form both homodimeric and heterodimeric complexes.ConclusionOur results expand the current knowledge of vertebrate CD8 dimerization and the interaction between CD8α with p/MHC I from an evolutionary perspective.

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“…With respect to cyprinid species, SVCV is the most widely spread viral pathogen [ 21 ]. SVCV has been shown to be a powerful inducer of ifn expression in carp cells [ 22 , 23 ]. Likewise, the up-regulation of ifn gene expression by SVCV infection has been found in zebrafish cells [ 24 , 25 , 26 ].…”

Section: Ssrna Virusesmentioning

confidence: 99%

“…Poly I:C consistently induces a rapid and strong interferon response in carp and zebrafish cells [ 22 , 23 , 25 , 26 , 29 , 77 ]. Mx RNA levels increased significantly following epithelioma papulosum cyprini (EPC) cells treatment either with poly I:C or with conditioned medium from cells treated with poly I:C [ 78 ].…”

Section: Dsrna Viruses and Poly I:cmentioning

confidence: 99%

Fish Innate Immune Response to Viral Infection—An Overview of Five Major Antiviral Genes

Ortega-Villaizán¹,

Chico²,

Pérez³

2022

Viruses

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Fish viral diseases represent a constant threat to aquaculture production. Thus, a better understanding of the cellular mechanisms involved in establishing an antiviral state associated with protection against virus replication and pathogenesis is paramount for a sustainable aquaculture industry. This review summarizes the current state of knowledge on five selected host innate immune-related genes in response to the most relevant viral pathogens in fish farming. Viruses have been classified as ssRNA, dsRNA, and dsDNA according to their genomes, in order to shed light on what those viruses may share in common and what response may be virus-specific, both in vitro (cell culture) as well as in vivo. Special emphasis has been put on trying to identify markers of resistance to viral pathogenesis. That is, those genes more often associated with protection against viral disease, a key issue bearing in mind potential applications into the aquaculture industry.

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Structural and Functional Analyses of Type I IFNa Shed Light Into Its Interaction With Multiple Receptors in Fish

Cited by 7 publications

References 47 publications

Transcriptome-wide analyses of early immune responses in lumpfish leukocytes upon stimulation with poly(I:C)

Transcriptome-wide analyses of early immune responses in lumpfish leukocytes upon stimulation with poly(I:C)

The first crystal structure of CD8αα from a cartilaginous fish

Fish Innate Immune Response to Viral Infection—An Overview of Five Major Antiviral Genes

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