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Developmental and Comparative Immunology
Pergamon Press
Developmental and Comparative Immunology

Pergamon Press

0145-305X

Developmental and Comparative Immunology/Journal Developmental and Comparative ImmunologySCIISTP
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    Identification of mRNA-miRNA-lncRNA regulatory network associated with the immune response to Aeromonas salmonicides infection in the black rockfish (Sebastes schlegelii)

    Gao, ChengbinCai, XinMa, LeLi, Chao...
    16页
    查看更多>>摘要：In aquaculture, Aeromonas salmonicides (A. salmonicida) is a main fish pathogen because of its nearly worldwide distribution, and broad host range. Recently, an increasing number of evidences have uncovered the roles of mRNA-miRNA-lncRNA network in fish diseases. In current study, RNA-seq was conducted in the black rockfish spleen following A. salmonicida infection at 0 h (Sp0 or control) and three different post-infection time-points (2 h: Sp2, 12 h: Sp12 and 24 h: Sp24, respectively) to comprehensively identify differentially expressed (DE) mRNAs, miRNAs and lncRNAs. Enrichment analysis and protein-protein interaction (PPI) analysis of DE mRNAs were performed. Then, expression and correlation analysis for mRNAs and their upstream miRNAs and lncRNAs were conducted. Finally, a total of 1364 mRNAs, 17 miRNAs and 1584 lncRNAs exhibited significantly differential expressions during bacterial infection in the black rockfish spleen. Functional enrichment analysis suggested that they were significantly enriched in several immune-related pathways, including Amino sugar and nucleotide sugar metabolism, Cell adhesion molecules (CAMs), Neuroactive ligand-receptor interaction, Nicotinate and nicotinamide metabolism, Pentose and glucuronate interconversions, Phagosome, Proteasome, etc. Subsequently, 1091 lncRNA-miRNA-mRNA pathways (323 in Sp2, 609 in Sp12 and 207 in Sp24) were constructed including 400 lncRNAs, 69 miRNAs, and 70 mRNAs. Meanwhile, NLRC3/novel-264/LNC_00116154 pathway demonstrated important immune modulating function in the black rockfish against A. salmonicida infection. Finally, the novel mRNA-miRNA-lncRNA sub-networks were established, among which all mRNAs and ncRNAs possessed significant predictive values for further studies for immune responses in the black rockfish.
      原文链接:
    • NSTL
    • Elsevier

    Transcriptome-wide analysis of cellular immune response stimulated by nuclear input of different down syndrome cell adhesion molecule intracellular domains

    Li, HaoZhao, YuehongZhang, XiaoliZhao, Hui...
    10页
    查看更多>>摘要：In arthropods, Dscam (Down syndrome cell adhesion molecule) produces multiple pathogen specific receptors via immune responsive alternative splicing, generating molecular complexity analogous to vertebrate antibodies. Fewer isoforms are produced by the exons encoding Dscam's intracellular domain (ICD); therefore, the present study aimed to determine the transcriptional response of Eriocheir sinensis to Dscam ICDs. In the group over expressing all cytoplasmic tail exons (ICD-FL), 1401 differentially expressed genes (DEGs) were identified; overexpressed of ICD constructs lacking exon-35 (ICD-delta 35) identified 413 DEGs; and overexpression of ICD constructs lacking exon-35 and exon-36 (ICD-delta 35 + 36) identified 22 DEGs. The DEGs were enriched in immunity and metabolism-related pathways. The expression of selected genes was confirmed using quantitative real-time reverse transcription PCR. The transcriptomes of Drosophila S2 cells overexpressing different ICDs were then determined. We identified key immune, metabolic, and cell proliferation-regulated genes and gene networks, providing insights into the membrane-to-nuclear signaling pathway of Dscam.
      原文链接:
    • NSTL
    • Elsevier

    Identification and function analysis of BmPxtA in the immune response regulated by PGE(2) of silkworm, Bombyx mori

    Shi, GuiqinZhou, YuanRen, Fei
    11页
    查看更多>>摘要：Prostaglandins (PGs) can mediate the immune response of insects to infection. Mammalian cyclooxygenase (COXs) is a key enzyme in the synthesis of PGs, and Pxt may be its homologous gene in some sequenced insect genomes. As a representative of Lepidoptera, the silkworm also contains PGs, but the biosynthetic source of PGs is still unclear. In this study, Sequence analysis showed that peroxinectin (BmPxtA) gene of silkworm was closely related to human COX gene, and its homologous protein had conserved domains corresponding to human COX. The expression of BmPxtA gene was the highest in the hemocytes and was induced by Nuclear Polyhedrosis Virus (NPV) challenge in the detected tissues. The quantitative polymerase chain reaction (qPCR) results showed that silencing BmPxtA mediated by RNA interference (RNAi) inhibited the expression of immune-related pathway genes, and specifically suppressed hemocyte-spreading and nodule formation in silkworm; Hemocyte-spreading and nodule formation were also inhibited by aspirin, a COX inhibitor. Treatment by PGE2 but not arachidonic acid (AA) rescued the immunosuppression; PGs concentrations was also inhibited by aspirin. PGE2, but not AA, treatment rescued the PGs concentrations. These results suggest that BmPxtA gene is associated with PG biosynthesis in silkworm and the immune response of silkworm was affected by regulating the concentrations of PGs.
      原文链接:
    • NSTL
    • Elsevier

    Effects of the interaction between a clip domain serine protease and a white spot syndrome virus protein on phenoloxidase activity

    Kwankaew, PattamapornMadsari, NaeemThongsoi, RatipornUtarabhand, Prapaporn...
    11页
    查看更多>>摘要：Clip domain serine proteinases participate in invertebrate innate immunity by acting as crucial enzymes in the signaling cascade involved in shrimp immunity. To functionally characterize its role in Fenneropenaeus mer-guiensis, FmclipSP cDNA was cloned and characterized. The FmclipSP gene comprised 1353 bp with an open reading frame of 1110 bp and encoded 369 amino acids. The protein contained clip and serine protease domains. FmClipSP mRNA is highly expressed in hemocytes, and its expression was significantly upregulated by bacterial or viral pathogen challenge. Furthermore, FmClipSP recombinant protein (rFmClipSP) was produced and possessed protease activity, stimulating prophenoloxidase activity. Additionally, rFmClipSP exhibited antibac-terial activity against pathogens and nonpathogens. ELISA results demonstrated the binding ability of rFmClipSP to a recombinant protein of VP28 (rVP28). Interestingly, the binding significantly inhibited prophenoloxidase activity. Altogether, we partially characterized the function of FmclipSP and demonstrated its association with VP28. This study indicates the importance of clipSP as a component of F. merguiensis innate immunity. However, the role of clipSP in crustaceans remains unclear and requires further investigation.
      原文链接:
    • NSTL
    • Elsevier

    microRNA-144 modulates the NF-?B pathway in miiuy croaker (Miichthys miiuy) by targeting I?Ba gene

    Yang, LiyuanZheng, WeiweiLv, XingXin, Shiying...
    6页
    查看更多>>摘要：MicroRNAs (miRNA) are non-coding RNAs that regulate many biochemical processes, such as cell growth, proliferation and immune responses. In this study, we investigated miR-144 as a regulator of I kappa B alpha that promotes the activation of NF-kappa B signaling pathway. And I kappa B alpha interact with p65 blocks nuclear translocation of NF-kappa B and anchors NF-kappa B in cytoplasmic quiescent cells in an inactive form. The seed region of miR-144 can regulate gene expression by binding to the 3' UTR of I kappa B alpha and repress I kappa B alpha expression at the post-transcriptional level. More importantly, miR-144 can promote the activation of p65 by inhibiting I kappa B alpha, thus affecting the NF-kappa B signaling pathway. Thus, preventing excessive inflammatory responses from causing autoimmune diseases will help to further understand the immunoregulatory mechanisms of miRNAs in fish after invasion by pathogens.
      原文链接:
    • NSTL
    • Elsevier

    Molecular characterization and immunoregulatory analysis of suppressors of cytokine signaling 1 (SOCS1) in black rockfish, Sebastes schlegeli

    Wang, GuanghuaLiu, WenqingWang, ChangbiaoWang, Jingjing...
    9页
    查看更多>>摘要：The suppressors of cytokine signaling (SOCS) family are important soluble mediators to inhibit signal transduction via the Janus kinase/signal transducer and activator of transcription (JAK-STAT) pathway in the innate and adaptive immune responses. SOCS1 is the primary regulator of a number of cytokines. In this study, two spliced transcripts of SOCS1 were identified and characterized from black rockfish (Sebastes schlegeli), named SsSOCS1a and SsSOCS1b. SsSOCS1a and SsSOCS1b contained conserved structural and functional domains including KIR region, ESS region, SH2 domain and SOCS box. SsSOCS1a and SsSOCS1b were distributed ubiquitously in all the detected tissues with the higher expression level in liver and spleen. After stimulation in vivo with Vibrio anguillarum and Edwardsiella tarda, the mRNA expression of SsSOCS1a and SsSOCS1b were induced in most of the immune-related tissues, including head kidney, spleen and liver. Meanwhile, poly I:C and IFN gamma up regulated the expression of SsSOCS1a and SsSOCS1b that reached the highest level at 24 h in macrophages in vitro. Luciferase assays in HEK293 cells showed SsSOCS1a and SsSOCS1b had the similar function in inhibiting ISRE activity after poly I:C and IFN gamma treatment. Furthermore, KIR domain in black rockfish was determined to have a negative regulatory role in IFN signaling. SsSOCS1a and SsSOCS1b were found to interact strongly with each other by Co-immunoprecipitation analyses. These results indicated that the function of SOCS1 in the negative regulation of IFN signaling is conserved from teleost to mammals which will be helpful to further understanding of the biological functions of teleosts SOCS1 in innate immunity.
      原文链接:
    • NSTL
    • Elsevier

    Modulation of fish immune response by interferon regulatory factor 4 in redlip mullet (Liza haematocheilus): Delineation through expression profiling, antiviral assay, and macrophage polarization analysis

    Harasgama, J. C.Kasthuriarachchi, T. D. W.Sirisena, D. M. K. P.Kwon, Hyukjae...
    7页
    查看更多>>摘要：Interferon regulatory factor 4 (IRF4) is a crucial member of IRF family, which acts as an imperative transcription factor in the development and maturation of multiple lineages of blood cells and also plays a pivotal role in host defense against microbial infections. In the present study, we aimed to investigate the detailed structural and functional aspects of a redlip mullet IRF4 homolog (LhIRF4). The LhIRF4 open reading frame consists of 1347 base pairs encoding 449 amino acids, with the DNA-binding domain sharing significant homology with that of other vertebrate IRF4 homologs. The highest transcription levels of LhIRF4 were observed in the mullet intestine and spleen under normal physiological conditions. Furthermore, a time-dependent upregulation of LhIRF4 transcription was observed in the spleen and head kidney tissues upon pathogenic challenges. When overexpressed in mullet cells, LhIRF4 was localized to the nucleus and significantly stimulated the transcription of several host antiviral genes. Moreover, the overexpression of LhIRF4 strongly inhibited the replication of viral hemorrhagic septicemia virus (VHSV) in vitro. The function of LhIRF4 in regulation of macrophage M2 polarization has also been evidently demonstrated in RAW 264.7 cells. Taken together, our findings indicate the profound role of LhIRF4 in modulating immune responses against microbial infections in redlip mullet.
      原文链接:
    • NSTL
    • Elsevier

    Molecular characterization of HEPCIDIN-1 (HAMP1) gene in red-bellied pacu (Piaractus brachypomus)

    Petano-Duque, Julieth MichelLozano-Villegas, Kelly JohannaCespedes-Rubio, Angel EnriqueRondon-Barragan, Iang Schroniltgen...
    7页
    查看更多>>摘要：Hepcidins are cysteine-rich peptides, which participate in iron metabolism regulation, the inflammatory and antimicrobial response. This study characterizes the hepcidin-1 (HAMP1) gene, its transcript expression in different tissues, as well as its regulation in a model of brain injury in Piaractus brachypomus. Bioinformatic analysis was carried out to determine conserved domains, glycosylation sites and protein structure of HAMP1, and probability that HAMP1 corresponds to an antimicrobial peptide (AMP). Relative gene expression of the P. brachypomus HAMP1 gene was determined by qPCR from cDNA of several tissues, a brain injury model, an organophosphate sublethal toxicity model and anesthetic experiment using the 2(-delta delta Ct) method. HAMP1 ORF encodes for a 91 aa pre-prohepcidin conformed for a prodomain with 42 aa and mature peptide of 25 aa. Mature domain was determined as an AMP. HAMP1 transcript is expressed in all the tissues, being higher in the spleen and liver. HAMP1 mRNA level was upregulated in the brain injury group, as well as in the olfactory bulb, optic chiasm and telencephalon of red-bellied pacu brain exposed to an organophosphate. In anesthetic experiment, HAMP1 mRNA level was upregulated in the liver and gills. HAMP1 gene of P. brachypomus may be involved in the inflammatory, antimicrobial, hypoxia and stress oxidative response.
      原文链接:
    • NSTL
    • Elsevier

    A flow cytometry based approach to identify distinct coelomocyte subsets of the purple sea urchin, Strongylocentrotus purpuratus

    Hudgell, Megan A. BarelaGrayfer, LeonSmith, L. Courtney
    8页
    查看更多>>摘要：The sea urchin, Strongylocentrotus purpuratus, possesses at least seven distinguishable cell populations in the coelomic fluid, which vary in morphology, size, and function. Of these, the large phagocytes, small phagocytes, and red spherule cells are thought to be key to the echinoid immune response. Because there are currently no effective and rapid means of evaluating sea urchin coelomocytes, we developed a flow cytometry based approach to identify these subsets from unseparated, unstained, live cells. In particular our gating strategy distinguishes between the large phagocytes, small phagocytes, red spherule cells, and a mixed population of vibratile cells and colorless spherule cells. This flow cytometry based analysis increases the speed and improves the reliability of coelomocyte analysis compared to differential cell counts by microscopy.
      原文链接:
    • NSTL
    • Elsevier

    Exosomes from H5N1 avian influenza virus-infected chickens regulate antiviral immune responses of chicken immune cells

    Hong, YeojinTruong, Anh DucVu, Thi HaoLee, Sooyeon...
    9页
    查看更多>>摘要：Exosomes (membrane-derived vesicles) enable intracellular communication by delivering lipids, proteins, DNA, and RNA from one cell to another. Highly pathogenic avian influenza virus (HPAIV) H5N1 causes considerable economic loss in the poultry industry and poses a public health concern. The host innate immune system defends against H5N1 infection by activating antiviral immune responses. This study aimed to demonstrated that immunomodulatory effects of exosomes from HPAIV H5N1-infected White Leghorn chickens on chicken macrophages, fibroblasts, T cell, and B cell lines. The expression of type I interferons (IFN-alpha and -beta) were highly upregulated in immune-related cell lines after treatment with exosomes derived from H5N1-infected chickens. Levels of pro-inflammatory cytokines, such as IFN-gamma, IL-1 beta, and CXCL8, were also elevated by the exosomes. The mitogen-activated protein kinase (MAPK) signaling pathway was stimulated in immune-related cells by such exosomes via phosphorylation of extracellular regulated kinases 1/2 and p38 signaling molecules. Furthermore, the H5N1 viral proteins, nucleoprotein (NP) and non-structural protein (NS1), were packaged in exosomes and successfully transferred to non-infected immune-related cells. Therefore, exosomes from H5N1-infected chickens induced pro-inflammatory cytokine expression and stimulated the MAPK signaling pathway by delivering key viral proteins. These findings would aid better understanding of the mechanism underlying the modulation of antiviral immune responses of host immune-related cells by viral-protein-carrying exosomes and support their further application as a novel exosome-based H5N1 AIV vaccine platform.
      原文链接:
    • NSTL
    • Elsevier