Developmental and comparative immunology最新文献

{"title":"Characterization of a fish-specific Immunoglobulin-like domain-containing protein (Igldcp) in zebrafish (Danio rerio) induced after nodavirus infection.","authors":"Nieves Martínez-López, Patricia Pereiro, Amaro Saco, Raquel Lama, Antonio Figueras, Beatriz Novoa","doi":"10.1016/j.dci.2024.105285","DOIUrl":"https://doi.org/10.1016/j.dci.2024.105285","url":null,"abstract":"<p><p>One of the most highly induced genes in zebrafish (Danio rerio) larvae after infection with the nodavirus red-spotted grouper nervous necrosis virus (RGNNV) was a member of the immunoglobulin superfamily (IgSF), which has remained uncharacterized and erroneously annotated in zebrafish and other fish species, such as galectin 17 (lgals17). We characterized this gene and named it immunoglobulin (Ig)-like domain-containing protein (igldcp), a new member of the IgSF that does not possess orthologs in mammals. Igldcp expression is induced by viral infection and it belongs to the group of interferon-stimulated genes (ISGs). In vitro overexpression of igldcp decreased RGNNV replication, whereas in vivo knockdown of this gene had the opposite effect, resulting in increased larval mortality. RNA-Seq analyses of larvae overexpressing igldcp in the absence or presence of infection with RGNNV showed that the main processes affected by Igldcp could be directly involved in the regulation of various cellular processes associated with the modulation of the immune system.</p>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142603423","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Molecular and functional characterization of short peptidoglycan recognition proteins in Vesicomyidae clam","authors":"Xue Kong ,&nbsp;Wei Wang ,&nbsp;Sunan Xia ,&nbsp;Ying Zhi ,&nbsp;Yuefeng Cai ,&nbsp;Haibin Zhang ,&nbsp;Xin Shen","doi":"10.1016/j.dci.2024.105284","DOIUrl":"10.1016/j.dci.2024.105284","url":null,"abstract":"<div><div>Within cold seep environments, the Vesicomyidae clam emerges as a prevalent species, distinguished by its symbiotic relationship with microorganisms housed within its organ gill. Given the extreme conditions and the symbiotic nature of this association, investigating the host's immune genes, particularly immune recognition receptors, is essential for understanding their role in facilitating host-symbiotic interactions. Three short peptidoglycan recognition proteins (PGRPs) were identified in the clam. AmPGRP-S1, -S2, and -S3 were found to possess conserved amidase binding sites and Zn²⁺ binding sites. Quantitative Real-time PCR (qRT-PCR) analysis revealed differential expression patterns among the PGRPs. AmPGRP-S1 and AmPGRP-S2 exhibited elevated expression levels in the gill, while AmPGRP-S3 displayed the highest expression in the adductor muscle. Functional experiments demonstrated that recombinant AmPGRP-S1, -S2, and -S3 (rAmPGRPs) exhibited binding capabilities to both L-PGN and D-PGN (peptidoglycan). Notably, rAmPGRP-S1 and -S2 possessed Zn²⁺-independent amidase activity, while rAmPGRP-S3 lacked this enzymatic function. rAmPGRPs were shown to bind to five different bacterial species. Among these, rAmPGRP-S1 inhibited <em>Escherichia coli</em> and <em>Bacillus subtilis</em>, while rAmPGRP-S2 and -S3 inhibited <em>Bacillus subtilis</em> in the absence of Zn^2<em>+</em>. In the presence of Zn²⁺, rAmPGRP-S1 and -S2 exhibited enhanced inhibitory activity against <em>Staphylococcus aureus</em> or <em>Bacillus subtilis</em>. These findings suggest that AmPGRPs may play a pivotal role in mediating the interaction between the host and endosymbiotic bacteria, functioning as PGN and microbe receptors, antibacterial effectors, and regulators of host-microbe symbiosis. These results contribute to our understanding of the adaptive mechanisms of deep-sea organisms to the challenging cold seep environments.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142567814","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Molecular depiction and functional delineation of E3 ubiquitin ligase MARCH5 in yellowtail clownfish (Amphiprion clarkii)","authors":"B.P.M. Vileka Jayamali ,&nbsp;H.M.S.M. Wijerathna ,&nbsp;D.M.K.P. Sirisena ,&nbsp;H.A.C.R. Hanchapola ,&nbsp;W.A.D.L.R. Warnakula ,&nbsp;U.P.E. Arachchi ,&nbsp;D.S. Liyanage ,&nbsp;Sumi Jung ,&nbsp;Qiang Wan ,&nbsp;Jehee Lee","doi":"10.1016/j.dci.2024.105283","DOIUrl":"10.1016/j.dci.2024.105283","url":null,"abstract":"<div><div>Membrane-associated Ring-CH 5 (MARCH5) is a mitochondrial E3 ubiquitin ligase playing a key role in the regulation of mitochondrial dynamics. In mammals, MARCH5 negatively regulates mitochondrial antiviral signaling (MAVS) protein aggregation during viral infection and hampers downstream type I interferon signaling to prevent excessive immune activation. However, its precise functional role in the teleost immune system remains unclear. This study investigated the molecular characteristics and immune response of the MARCH5 ortholog in <em>Amphiprion clarkii</em> (<em>A. clarkii</em>; AcMARCH5). The predicted AcMARCH5 protein sequence consists of 287 amino acids with a molecular weight of 32.02 kDa and a theoretical isoelectric point of 9.11. It contains four C-terminal transmembrane (TM) domains and an N-terminal RING cysteine-histidine (CH) domain, which directly regulates ubiquitin transfer. Multiple sequence alignment revealed a high level of conservation between AcMARCH5 and its orthologs in other vertebrate species. Under normal physiological conditions, <em>AcMARCH5</em> showed the highest mRNA expression in the muscle, brain, and kidney tissues of <em>A. clarkii</em>. Upon stimulation with polyinosinic:polycytidylic acid (Poly I:C), lipopolysaccharide (LPS), and <em>Vibrio harveyi</em>, <em>AcMARCH5</em> expression was drastically modulated. Functional assays showed that overexpression of AcMARCH5 in fathead minnow (FHM) cells downregulated antiviral gene expression, accompanied by enhanced viral hemorrhagic septicemia virus (VHSV) replication. In murine macrophages, AcMARCH5 overexpression markedly reduced the production of pro-inflammatory cytokines in response to poly I:C treatment. Additionally, AcMARCH5 exhibited an anti-apoptotic effect in H₂O₂-treated FHM cells. Collectively, these results suggest that AcMARCH5 may play a role in maintaining cellular homeostasis under disease and stress conditions in <em>A. clarkii</em>.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142557392","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dysregulated proinflammatory cytokines and immune-related miRNAs in ASK cells exposed to 17⍺-Ethynyl estradiol and 4-nonylphenol","authors":"Carolina Salazar ,&nbsp;Nicolás Ojeda ,&nbsp;Luis Mercado","doi":"10.1016/j.dci.2024.105282","DOIUrl":"10.1016/j.dci.2024.105282","url":null,"abstract":"<div><div>Endocrine Disruptor Compounds (EDCs) in the aquatic environment have acquired pronounced relevance due to their toxic effect on the aquatic flora and fauna. Xenoestrogens are EDCs that possess estrogenic activity and, thus, disrupt normal estrogen signaling, affecting different functions, such as immune system processes. Two relevant xenoestrogens discarded into fresh and seawater are 4-nonylphenol (NP) and 17⍺-Ethynyl Estradiol (EE2). Considering that the piscicultures of <em>Salmo salar</em> can be located at sites of potential exposure to xenoestrogen-containing effluxes, it is crucial to understand the effect of xenoestrogens on the immune response and its possible molecular mechanism in this species. Our studies reveal an increase in the expression of the receptor <em>era</em> and <em>erb</em> at early times of exposure, a disrupted expression of pro-inflammatory cytokines (<em>il1b</em> and <em>tnfa</em>), an upregulation of ssa-miR-146a-5p, ssa-miR-125 b-5p, and downregulation of ssa-miR-145–5p in ASK cells exposed to estrogen and xenoestrogen, could potentially lead to new strategies for mitigating the effects of xenoestrogens on <em>Salmo salar</em> immune response.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-10-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142496903","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"CqProfilin enhances WSSV infection by promoting viral intracellular transport through binding to both viral nucleocapsid and actin cytoskeleton","authors":"Dong-li Li ,&nbsp;Wen-lin Wu ,&nbsp;Hai-peng Liu","doi":"10.1016/j.dci.2024.105281","DOIUrl":"10.1016/j.dci.2024.105281","url":null,"abstract":"<div><div>White spot syndrome virus (WSSV) is a large nuclear-replicating DNA virus of crustaceans such as shrimp and crayfish; however, the molecular mechanisms facilitating its transport from the invasion site to the cell nucleus have not yet been well elucidated. In this study, a <em>Cq</em>Profilin (<em>Cq</em>PFN) with a conserved PROF domain was identified from the red claw crayfish <em>Cherax quadricarinatus</em>. <em>CqPFN</em> was ubiquitously expressed in all examined tissues and hemocyte, with the highest levels in the hemocyte, followed by hematopoietic tissue (Hpt) from which the hemocyte were derived in crayfish. The transcript of WSSV genes such as <em>IE1</em> and <em>VP28</em> was obviously decreased both <em>in vivo</em> in hemocyte and Hpt, as well as <em>in vitro</em> in cultured Hpt cells, after <em>CqPFN</em> gene silencing; in contrast, the expression of viral genes was significantly increased by the introduction of a recombinant <em>Cq</em>PFN protein in Hpt cells <em>in vitro</em>. Moreover, <em>Cq</em>PFN was clearly colocalized with the main viral nucleocapsid protein VP664 and F-actin cytoskeleton, respectively, during the early stage of WSSV infection in Hpt cells. In addition, <em>Cq</em>PFN was confirmed to interact with a truncated VP664^2,405-2,535 and another viral nucleocapsid protein VP15 of WSSV and <em>Cq</em>β-Actin from Hpt by co-immunoprecipitation assays. Further studies found that VP664 also colocalized with F-actin in the Hpt cell cytoplasm after WSSV infection, suggesting that the actin cytoskeleton was involved in the intracellular transport of incoming viral nucleocapsid. Taken together, <em>Cq</em>PFN might combine with the actin cytoskeleton to promote WSSV infection through binding with viral nucleocapsid proteins VP664 and VP15, promoting intracellular transport of viral incoming nucleocapsid for further releasing genome into the nucleus for transcription. Collectively, these results provided an understanding of the WSSV pathogenesis, which will contribute to the development of an antiviral strategy against WSSV disease.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142460364","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dynamic alterations in T-lymphocyte subsets assessed by flow cytometry in chickens following exposure to infectious bursal disease virus: A systematic review","authors":"Sara Pedrazzoli,&nbsp;Giulia Graziosi,&nbsp;Roberta Salaroli,&nbsp;Elena Catelli,&nbsp;Caterina Lupini","doi":"10.1016/j.dci.2024.105280","DOIUrl":"10.1016/j.dci.2024.105280","url":null,"abstract":"<div><div>Infectious bursal disease virus (IBDV) is a significant pathogen in poultry, causing acute immunosuppressive disease in young chickens. While B-lymphocyte involvement in IBDV pathogenesis is known, the role of T-cells is incompletely understood. This systematic review presents the alterations in chicken T-lymphocyte subsets after IBDV exposure, assessed by flow cytometry analysis. Four databases were queried for identifying eligible studies focused on experimental infections measuring T-lymphocyte changes in the bursa of Fabricius, spleen, thymus, and peripheral blood mononuclear cells. Of 488 studies found, 25 met the pre-established criteria and were included in the qualitative synthesis of results. Most studies analysed T-lymphocyte responses during the acute phase of IBDV infection, primarily focusing on CD4⁺ and CD8⁺ T-cells. Other subsets, such as γδ T-cells and double-positive CD4⁺CD8⁺ T-cells, were less frequently investigated. An increase in T-lymphocytes was noted in the bursa of Fabricius, suggesting their active role in viral clearance. In the spleen, CD4⁺ T-cells commonly increased, while CD8⁺ responses varied among studies. Increased levels in T-cells were also noted during the chronic infection in the bursa of Fabricius, possibly due to persistent viral antigens. Overall, variations in flow cytometry methods and T-cell output reporting were noted among studies.</div><div>Based on the data collected, further investigation into diverse T-cell subpopulations beyond CD4⁺ and CD8⁺ is needed, as well as the standardization of flow cytometry assays in chickens.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142460365","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Unveiling the immunomodulatory role of soluble chicken fractalkine: Insights from functional characterization and pathway activation analyses","authors":"Thi Hao Vu ,&nbsp;Chaeeun Kim ,&nbsp;Anh Duc Truong ,&nbsp;Hyun S. Lillehoj ,&nbsp;Yeong Ho Hong","doi":"10.1016/j.dci.2024.105279","DOIUrl":"10.1016/j.dci.2024.105279","url":null,"abstract":"<div><div>This study describes the first successful cloning and functional characterization of chicken CX3CL1, a chemokine involved in immune cell migration and inflammatory responses. Evolutionary analyses revealed its close relation to CX3CL1 from other avian species, particularly duck, turkey, and quail. Structurally, chicken CX3CL1 includes a signal peptide and a chemokine interleukin-8-like domain characterized by unique alpha-helices and disulfide bonds. Additionally, we produced and purified recombinant CX3CL1 protein and assessed its endotoxin levels. Chemotaxis assays revealed that CX3CL1 significantly enhances the migration of HD11 macrophages and CU91 T cells. Furthermore, recombinant CX3CL1 induced the expression of pro-inflammatory cytokines (TNF-α, IFN-β, IFN-γ, IL-6, and CCL20) in a time-dependent manner, while exerting differential effects on anti-inflammatory cytokines (IL-4, IL-10). Conversely, transfection with siCX3CL1 or siCX3CR1 led to the downregulation of these responses. We also observed activation of the MAPK, NF-κB, and JAK/STAT pathways, evidenced by increased phosphorylation of key signaling molecules. These findings underscore the crucial role of chicken CX3CL1 in regulating immune responses, cell migration, and the activation of key signaling pathways. This study provides valuable insights into the immunomodulatory functions of soluble CX3CL1, highlighting its potential as a therapeutic target for inflammatory conditions and enhancing our understanding of immune cell dynamics.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142444729","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The essential function of cathepsin X of the orange-spotted grouper, Epinephelus coioides during SGIV infection","authors":"Yuexuan Wang ,&nbsp;Yewen Wang ,&nbsp;Yunxiang Jiang ,&nbsp;Qiwei Qin ,&nbsp;Shina Wei","doi":"10.1016/j.dci.2024.105278","DOIUrl":"10.1016/j.dci.2024.105278","url":null,"abstract":"<div><div>Cathepsin X, a class of cysteine proteases in the lysosome, involved in intracellular protein degradation processes. Numerous reports revealed that many kinds of cysteine proteases played a crucial role in pathogen invasion. To investigate the relationship between cathepsin X of teleost fish and virus infection, EcCX was cloned and characterized in the orange-spotted grouper, <em>Epinephelus coioides</em>. The open reading frame (ORF) of EcCX included 909 nucleotides and encoded a protein consisting of 302 amino acids, which shared 75% and 56% identity with zebrafish and humans, respectively. The protein EcCX mainly consisted of a signal peptide (1–19 aa), a pro-pre-peptide region (20–55 aa), and a mature cysteine protease region (56–302 aa). Subcellular localization analysis showed that EcCX was mainly distributed in the cytoplasm, but EcCX ectoped to the vicinity of apoptotic vesicles in FHM cells during SGIV infection. Following stimulation with SGIV or Poly (dA:dT), there was a notable rise in the expression levels of EcCX. EcCX overexpression facilitated virus infection, upregulated the production of inflammatory factors, and induced the activation of the NF-κB promoter. Furthermore, the overexpression of EcCX also accelerated the process of SGIV-induced apoptosis, potentially by enhancing the promoter activity of P53 and AP-1. Overall, our findings demonstrated a correlation between the function of EcCX and SGIV infection, providing a new understanding of the mechanisms involved in fish virus infection.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142444730","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transcriptomic and microbiome analyses of copepod Apocyclops royi in response to an AHPND-causing strain of Vibrio parahaemolyticus","authors":"Natkanokporn Prayoonmaneerat ,&nbsp;Walaiporn Charoensapsri ,&nbsp;Piti Amparyup ,&nbsp;Chanprapa Imjongjirak","doi":"10.1016/j.dci.2024.105277","DOIUrl":"10.1016/j.dci.2024.105277","url":null,"abstract":"<div><div>Copepods are small crustaceans that live in microorganism-rich aquatic environments and provide a key supply of live food for fish and shellfish larviculture. To better understand the host-pathogen interaction between the copepod and <em>Vibrio parahaemolyticus</em> causing acute hepatopancreatic necrosis disease (VP_AHPND), the comparative transcriptome and microbiome analyses were conducted in copepod <em>Apocyclops royi</em>-TH following VP_AHPND infection. Transcriptome analysis identified a total of 836 differentially expressed genes, with 275 upregulated and 561 downregulated genes. Subsequent analysis showed that a total of 37 differentially expressed genes were associated with the innate immune system, including 16 upregulated genes related to Toll-like receptor signaling pathway, antimicrobial peptides, and stress response genes, and 21 downregulated genes associated with immunological modulators, signaling molecules, and apoptosis-related proteins. Analysis of the copepod microbiome following VP_AHPND infection showed that the microbes changed significantly after bacterial infection, with a reduced alpha diversity accompanied by the increased level of Proteobacteria and decreased levels of Bdellovibrionota, Bacteroidota, and Verrucomicrobiota. The population of <em>Vibrio</em> genera were increased significantly, while several other genera, including <em>Denitromonas</em>, <em>Nitrosomonas</em>, <em>Blastopirellula</em>, <em>Fusibacter</em>, <em>Alteromonas</em>, <em>KI89A_clade</em>, and <em>Ruegeria</em>, were decreased significantly after infection. These findings suggest that VP_AHPND infection has a significant impact on the immune defense and the composition of the copepod microbiota.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142343554","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification and antibacterial activity of a novel phage-type lysozyme from the freshwater mussel Hyriopsis cumingii","authors":"Hui Xu ,&nbsp;Wenjuan Dai ,&nbsp;Zhengyu Xiong ,&nbsp;NaNa Huang ,&nbsp;Yanrui Wang ,&nbsp;Zhe Yang ,&nbsp;Shanshan Luo ,&nbsp;Jielian Wu","doi":"10.1016/j.dci.2024.105272","DOIUrl":"10.1016/j.dci.2024.105272","url":null,"abstract":"<div><div>A cDNA encoding a phage-type lysozyme, designated as HcPLYZ, was successfully cloned from <em>Hyriopsis cumingii</em>. The full-length cDNA sequence of HcPLYZ was determined to be 896 base pairs in length. Analysis revealed the absence of a signal peptide at its N-terminus, and identified two highly conserved phage-type lysozyme activity sites, Glu²⁰ and Asp²⁹, within the deduced amino acid sequence of HcPLYZ. The results of the cloning and sequencing symbiotic bacteria in tissues were consistent with those obtained using tissue cDNA as the template, suggesting that HcPLYZ may originate a symbiotic bacterium. The expression levels of HcPLYZ mRNA exhibited significant variations across different tissues. Successful expression was induced using IPTG, and the native recombinant protein was subsequently purified through affinity chromatography employing Ni²⁺, and the optimal pH and temperature of which were determined to be 5.5 and 50 °C, respectively. Following exposure to <em>Aeromonas hydrophila</em>, there was a significant increase in the levels of HcPLYZ mRNA in the hemocytes, hepatopancreas, and gills. HcPLYZ was demonstrated the inhibition activity of 55% and 83% against <em>Micrococcus lysodeikticus</em> under pH 5.5 and 50 °C conditions, respectively. These results suggested that HcPLYZ possessed antibacterial activity against both <em>A. hydrophila</em> and <em>M. lysodeikticus</em>.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142343552","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}