Crustaceans, like other invertebrates, lack adaptive immunity and rely primarily on innate immune responses for defense against invading pathogens. Cytokines are signaling molecules that play vital roles in coordinating innate immune responses. Cytokine-like molecules such as astakines, interferon-like proteins (namely Vago), Spätzle, interleukin-like proteins, and tumor necrosis factors have been identified in crustaceans including shrimp, crayfish, and crabs. These molecules, although structurally distinct from vertebrate cytokines, have critical roles in regulating innate immune responses. They mediate processes such as hemocyte proliferation, production of antimicrobial peptides, antiviral response, and anti-inflammation through key immune signaling pathways including Toll, JAK/STAT, and IMD. Astakines are particularly important in crustacean hematopoiesis; Vago is involved in antiviral immunity, and Spätzle functions as a key ligand in Toll signaling. Interleukin-like proteins and tumor necrosis factors, although less well characterized in crustaceans, likely play crucial roles in the innate immune system. Studying the functions of these cytokine-like proteins enables better understanding crustacean immunity and has significant implications for improving disease management in aquaculture.
{"title":"Functional insights into cytokine-like molecules in crustacean innate immunity","authors":"Warumporn Yingsunthonwattana, Zittipong Nanakorn, Anchalee Tassanakajon","doi":"10.1016/j.dci.2025.105528","DOIUrl":"10.1016/j.dci.2025.105528","url":null,"abstract":"<div><div>Crustaceans, like other invertebrates, lack adaptive immunity and rely primarily on innate immune responses for defense against invading pathogens. Cytokines are signaling molecules that play vital roles in coordinating innate immune responses. Cytokine-like molecules such as astakines, interferon-like proteins (namely Vago), Spätzle, interleukin-like proteins, and tumor necrosis factors have been identified in crustaceans including shrimp, crayfish, and crabs. These molecules, although structurally distinct from vertebrate cytokines, have critical roles in regulating innate immune responses. They mediate processes such as hemocyte proliferation, production of antimicrobial peptides, antiviral response, and anti-inflammation through key immune signaling pathways including Toll, JAK/STAT, and IMD. Astakines are particularly important in crustacean hematopoiesis; Vago is involved in antiviral immunity, and Spätzle functions as a key ligand in Toll signaling. Interleukin-like proteins and tumor necrosis factors, although less well characterized in crustaceans, likely play crucial roles in the innate immune system. Studying the functions of these cytokine-like proteins enables better understanding crustacean immunity and has significant implications for improving disease management in aquaculture.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"173 ","pages":"Article 105528"},"PeriodicalIF":2.4,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145582049","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}
Pub Date : 2025-12-01DOI: 10.1016/j.dci.2025.105529
Zhitao Qi , Jinquan He , Chenfei Gu , Yushuai Xie , Mingzhu Pan , Qihuan Zhang , Youchuan Wei
CD28 signaling pathway plays critical roles in the T cell activation. The golden pompano (Trachinotus ovatus, Tro) is a commercially important marine fish that widely cultured in southern China. Frequently pathogenic disease outbreaks greatly hinder the industry of golden pompano. Understanding the T cell activation signaling pathway of golden pompano contributes to prevent pathogenic diseases using immunological strategies. In the present study, three molecules of CD28 signaling pathway (TroCD28, growth factor receptor-bound protein 2 (TroGrb2) and SH2 domain-containing leukocyte phosphoprotein of 76 kDa (TroSLP-76)) were identified from golden pompano. The cDNA of the three genes respectively encoded 248, 217 and 375 amino acids. TroCD28, TroGrb2 and TroSLP-76 contained several conserved functional domains, similar to their mammalian counterparts. qPCR analysis revealed that these three genes were ubiquitously expressed in all selected tissues and peripheral blood lymphocytes (PBLs), with high expressions in immune-related tissues (spleen, head kidney, gill). Further, expressions of these three genes in head kidney were significantly induced at different time points post lipopolysaccharide (LPS) or polyriboinosinic polyribocytidylic acid (polyI:C) stimulation and Vibrio alginolyticus infection. The coimmunoprecipitation (Co-IP) assay verified the TroCD28-TroGrb2 interaction. These results provide basis for understanding the process of T cell activation in golden pompano.
{"title":"Characterization of the CD28 signaling pathway related genes (CD28, Grb2 and SLP-76) of golden pompano (Trachinotus ovatus)","authors":"Zhitao Qi , Jinquan He , Chenfei Gu , Yushuai Xie , Mingzhu Pan , Qihuan Zhang , Youchuan Wei","doi":"10.1016/j.dci.2025.105529","DOIUrl":"10.1016/j.dci.2025.105529","url":null,"abstract":"<div><div>CD28 signaling pathway plays critical roles in the T cell activation. The golden pompano (<em>Trachinotus ovatus</em>, Tro) is a commercially important marine fish that widely cultured in southern China. Frequently pathogenic disease outbreaks greatly hinder the industry of golden pompano. Understanding the T cell activation signaling pathway of golden pompano contributes to prevent pathogenic diseases using immunological strategies. In the present study, three molecules of CD28 signaling pathway (TroCD28, growth factor receptor-bound protein 2 (TroGrb2) and SH2 domain-containing leukocyte phosphoprotein of 76 kDa (TroSLP-76)) were identified from golden pompano. The cDNA of the three genes respectively encoded 248, 217 and 375 amino acids. TroCD28, TroGrb2 and TroSLP-76 contained several conserved functional domains, similar to their mammalian counterparts. qPCR analysis revealed that these three genes were ubiquitously expressed in all selected tissues and peripheral blood lymphocytes (PBLs), with high expressions in immune-related tissues (spleen, head kidney, gill). Further, expressions of these three genes in head kidney were significantly induced at different time points post lipopolysaccharide (LPS) or polyriboinosinic polyribocytidylic acid (polyI:C) stimulation and <em>Vibrio alginolyticus</em> infection. The coimmunoprecipitation (Co-IP) assay verified the TroCD28-TroGrb2 interaction. These results provide basis for understanding the process of T cell activation in golden pompano.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"173 ","pages":"Article 105529"},"PeriodicalIF":2.4,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145615645","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}
Pub Date : 2025-12-01DOI: 10.1016/j.dci.2025.105531
Hayley Brown , Selma Schmidt , Michelle Thom , Adam Kuča , Claudine Porta , John A. Hammond , Marie Bonnet-Di Placido , Wilhelm Gerner
With their capacity for antibody production, B cells underpin many protective immune responses, but they can also contribute to immunopathology and autoimmunity. Studying their differentiation following antigen encounter is therefore a key aspect of fundamental immunological research. However, in many livestock species, including cattle, the study of B cell subsets is limited by the restricted availability of immunological tools. Here, we demonstrate that cross-reacting anti-human and anti-murine antibodies used in combination with established anti-bovine reagents can be used to identify three major cattle B cell subsets by flow cytometry: a) total mature B cells, b) plasma cells and plasmablasts, and c) germinal centre (GC) B cells. For this, we employed antibodies targeting CD79α, Pax5, Blimp1, IRF4, and Bcl6. CD79α/Pax5 co-expressing total B cells largely overlapped with the previously described CD14−CD40+ phenotype. Putative plasma cells, defined by Blimp1/IRF4 co-expression, were detected in blood and lymph nodes and exhibited reduced Pax5 expression, consistent with a plasma cell phenotype. In contrast, putative GC B cells, identified by Bcl6 expression, were only observed in lymph nodes, as expected for this population. Using foot-and-mouth disease virus-like particles (VLPs) as probes to detect antigen-specific B cells in blood and lymph nodes from VLP-immunised cattle, we show that our newly established phenotypes facilitate the dissection of antigen-driven B cell responses. Notably, VLP-binding B cell subsets were predominantly detected in lymph node cell preparations draining the site of vaccination. Thus, this work significantly expands the toolbox available for studying B cell biology in cattle.
{"title":"Transcription factor expression for identifying B cell subsets in cattle by flow cytometry","authors":"Hayley Brown , Selma Schmidt , Michelle Thom , Adam Kuča , Claudine Porta , John A. Hammond , Marie Bonnet-Di Placido , Wilhelm Gerner","doi":"10.1016/j.dci.2025.105531","DOIUrl":"10.1016/j.dci.2025.105531","url":null,"abstract":"<div><div>With their capacity for antibody production, B cells underpin many protective immune responses, but they can also contribute to immunopathology and autoimmunity. Studying their differentiation following antigen encounter is therefore a key aspect of fundamental immunological research. However, in many livestock species, including cattle, the study of B cell subsets is limited by the restricted availability of immunological tools. Here, we demonstrate that cross-reacting anti-human and anti-murine antibodies used in combination with established anti-bovine reagents can be used to identify three major cattle B cell subsets by flow cytometry: a) total mature B cells, b) plasma cells and plasmablasts, and c) germinal centre (GC) B cells. For this, we employed antibodies targeting CD79α, Pax5, Blimp1, IRF4, and Bcl6. CD79α/Pax5 co-expressing total B cells largely overlapped with the previously described CD14<sup>−</sup>CD40<sup>+</sup> phenotype. Putative plasma cells, defined by Blimp1/IRF4 co-expression, were detected in blood and lymph nodes and exhibited reduced Pax5 expression, consistent with a plasma cell phenotype. In contrast, putative GC B cells, identified by Bcl6 expression, were only observed in lymph nodes, as expected for this population. Using foot-and-mouth disease virus-like particles (VLPs) as probes to detect antigen-specific B cells in blood and lymph nodes from VLP-immunised cattle, we show that our newly established phenotypes facilitate the dissection of antigen-driven B cell responses. Notably, VLP-binding B cell subsets were predominantly detected in lymph node cell preparations draining the site of vaccination. Thus, this work significantly expands the toolbox available for studying B cell biology in cattle.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"173 ","pages":"Article 105531"},"PeriodicalIF":2.4,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145615641","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}
Pub Date : 2025-11-30DOI: 10.1016/j.dci.2025.105533
Shanjie Zha , Siyi Huang , Luxia Zhang , Xinguo Zhao , Hongxing Liu , Wei Shi , Yongbo Bao
Exosomes are crucial for intercellular immune communication, but their function in marine bivalve immunity remains poorly understood. This study investigated the immunomodulatory role of hemocyte-derived exosomes in the blood clam Anadara granosa during Vibrio parahaemolyticus challenge. Exosomes isolated from hemolymph displayed typical cup-shaped morphology (149–153 nm diameter) and expressed canonical markers (CD63, TSG101). Functional analysis demonstrated that exosomes from Vibrio-challenged clams significantly enhanced hemocyte phagocytosis and reduced bacterial loads. To uncover the underlying mechanism, small RNA sequencing of the exosomal cargo identified 107 differentially expressed miRNAs (56 upregulated, 51 downregulated) upon bacterial challenge. Bioinformatic analysis revealed that these miRNAs primarily target genes involved in endocytosis, autophagy, and ubiquitin-mediated proteolysis. Notably, quantitative PCR validation confirmed the differential expression of six key miRNAs involved in phagocytosis regulation. These findings provide the first evidence that A. granosa hemocytes utilize exosomal miRNAs to regulate antibacterial response by enhanced phagocytosis. The identified exosomal miRNAs represent promising biomarkers for immune assessment and potential targets for developing RNA-based therapeutics to enhance disease resistance aquaculture.
{"title":"Exosomal miRNA-mediated enhancement of hemocyte immunity in Vibrio-stressed blood clam (Anadara granosa)","authors":"Shanjie Zha , Siyi Huang , Luxia Zhang , Xinguo Zhao , Hongxing Liu , Wei Shi , Yongbo Bao","doi":"10.1016/j.dci.2025.105533","DOIUrl":"10.1016/j.dci.2025.105533","url":null,"abstract":"<div><div>Exosomes are crucial for intercellular immune communication, but their function in marine bivalve immunity remains poorly understood. This study investigated the immunomodulatory role of hemocyte-derived exosomes in the blood clam <em>Anadara granosa</em> during <em>Vibrio parahaemolyticus</em> challenge. Exosomes isolated from hemolymph displayed typical cup-shaped morphology (149–153 nm diameter) and expressed canonical markers (CD63, TSG101). Functional analysis demonstrated that exosomes from <em>Vibrio</em>-challenged clams significantly enhanced hemocyte phagocytosis and reduced bacterial loads. To uncover the underlying mechanism, small RNA sequencing of the exosomal cargo identified 107 differentially expressed miRNAs (56 upregulated, 51 downregulated) upon bacterial challenge. Bioinformatic analysis revealed that these miRNAs primarily target genes involved in endocytosis, autophagy, and ubiquitin-mediated proteolysis. Notably, quantitative PCR validation confirmed the differential expression of six key miRNAs involved in phagocytosis regulation. These findings provide the first evidence that <em>A. granosa</em> hemocytes utilize exosomal miRNAs to regulate antibacterial response by enhanced phagocytosis. The identified exosomal miRNAs represent promising biomarkers for immune assessment and potential targets for developing RNA-based therapeutics to enhance disease resistance aquaculture.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"174 ","pages":"Article 105533"},"PeriodicalIF":2.4,"publicationDate":"2025-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145660557","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}
Pub Date : 2025-11-20DOI: 10.1016/j.dci.2025.105527
Xinyue Ren , Zhou Qin , Renhui Liu , Ting Chen , Tiehao Lin , Xinyu Sun , Jiasheng Huang , Wenjie Pan , Caixia Long , Chunhua Ren , Haipeng Qin , Peiming Zheng , Aifen Yan , Lihong Yuan
To date, the understanding of lectin-related immunity in sea cucumbers remains limited. Through a comprehensive genome-wide analysis, 133 lectin genes were identified from Holothuria leucospilota, a sea cucumber species with high ecological and economic significance in the Pacific and Indian Oceans. These lectins were classified into five subtypes, with C-type lectin being the predominant form, and the presence of SUEL-like lectin and Malectin reported here for the first time in sea cucumbers. Transcriptomic analysis across twelve tissues and sixteen developmental stages revealed ubiquitous expression of H. leucospilota lectins. A key finding was the compartmentalization of lectin systems: SUEL-like lectins expressed in the body wall constitute an “external” defense system, while C-type lectins produced by coelomocytes form an “internal” immune system. This spatial and functional specialization reflects a sophisticated immune strategy in invertebrates, enabling targeted pathogen recognition at different bodily interfaces. Furthermore, distinct immune responses to Vibrio, LPS, and poly(I:C) were observed among different lectins. We also identified an ovary-specific C-type lectin, suggesting a potential role in early embryonic development. This study not only fills a knowledge gap regarding lectin genes in sea cucumbers but also establishes a broader framework for understanding complex immune mechanisms in echinoderms.
{"title":"Genome-wide analysis unveils the differentiated “external” and “internal” lectin system in the tropical sea cucumber Holothuria leucospilota","authors":"Xinyue Ren , Zhou Qin , Renhui Liu , Ting Chen , Tiehao Lin , Xinyu Sun , Jiasheng Huang , Wenjie Pan , Caixia Long , Chunhua Ren , Haipeng Qin , Peiming Zheng , Aifen Yan , Lihong Yuan","doi":"10.1016/j.dci.2025.105527","DOIUrl":"10.1016/j.dci.2025.105527","url":null,"abstract":"<div><div>To date, the understanding of lectin-related immunity in sea cucumbers remains limited. Through a comprehensive genome-wide analysis, 133 lectin genes were identified from <em>Holothuria leucospilota</em>, a sea cucumber species with high ecological and economic significance in the Pacific and Indian Oceans. These lectins were classified into five subtypes, with C-type lectin being the predominant form, and the presence of SUEL-like lectin and Malectin reported here for the first time in sea cucumbers. Transcriptomic analysis across twelve tissues and sixteen developmental stages revealed ubiquitous expression of <em>H. leucospilota</em> lectins. A key finding was the compartmentalization of lectin systems: SUEL-like lectins expressed in the body wall constitute an “external” defense system, while C-type lectins produced by coelomocytes form an “internal” immune system. This spatial and functional specialization reflects a sophisticated immune strategy in invertebrates, enabling targeted pathogen recognition at different bodily interfaces. Furthermore, distinct immune responses to <em>Vibrio</em>, LPS, and poly(I:C) were observed among different lectins. We also identified an ovary-specific C-type lectin, suggesting a potential role in early embryonic development. This study not only fills a knowledge gap regarding lectin genes in sea cucumbers but also establishes a broader framework for understanding complex immune mechanisms in echinoderms.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"173 ","pages":"Article 105527"},"PeriodicalIF":2.4,"publicationDate":"2025-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145570338","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}
Pub Date : 2025-11-18DOI: 10.1016/j.dci.2025.105526
Bernard Yi Zhe Thian , Mohd Nasir Nurul Fatimah , Abdul Rahman Omar , Huzlinda Hussin , Hui Kian Ong , Chuan Loo Wong , Abdul Razak Mariatulqabtiah , Kok Lian Ho , Wen Siang Tan
H9N2 and H5N2 avian influenza A viruses (IAVs) are two of the most widespread and economically damaging subtypes affecting the poultry industry. Controlling these viruses in poultry remains challenging, highlighting the need for a vaccine that can confer broad and effective protection across multiple avian IAV subtypes. Previously, we fused two copies of the matrix 2 ectodomain (M2e) of human and avian IAV (2xhuM2e-2xavM2e) at the C-terminus of Macrobrachium rosenbergii nodavirus capsid protein (NvC). This chimeric protein (NvC-2xhuM2e-2xavM2e) self-assembled into virus-like particles (VLPs), which elicited broad immune responses and activated humoral and cellular immune responses in chickens. In this study, we evaluated the protective efficacy of these chimeric VLPs in White Leghorn chickens against H9N2 and H5N2 infections. Chickens immunized with the chimeric VLPs produced a significant level of M2e antibodies that reacted with the M2e of H9N2 and H5N2 IAVs. Following viral infections, the chimeric VLPs mitigated weight loss in H9N2-infected chickens and alleviated delayed weight gain in H5N2-infected chickens. Furthermore, the chimeric VLPs reduced the clinical signs in immunized chickens challenged with either H9N2 or H5N2 avian IAV. Viral loads in the lung tissues of the immunized chickens were significantly lower compared to the negative control group. The chimeric VLPs also enhanced H9N2 and H5N2 viral clearance, showing a significant reduction in viral shedding in the oropharynges and cloacae of the immunized chickens. Furthermore, the VLPs activated both inflammatory and anti-inflammatory cytokines, which were associated with enhanced viral clearance while reducing lung and tracheal immunopathology. Collectively, these findings demonstrate the potential of the chimeric VLPs as a cross-protective vaccine against both H9N2 and H5N2 infections in poultry.
{"title":"Virus-like particles of Macrobrachium rosenbergii nodavirus displaying M2e of influenza A viruses protect White Leghorn chickens against heterologous H9N2 and H5N2 challenges","authors":"Bernard Yi Zhe Thian , Mohd Nasir Nurul Fatimah , Abdul Rahman Omar , Huzlinda Hussin , Hui Kian Ong , Chuan Loo Wong , Abdul Razak Mariatulqabtiah , Kok Lian Ho , Wen Siang Tan","doi":"10.1016/j.dci.2025.105526","DOIUrl":"10.1016/j.dci.2025.105526","url":null,"abstract":"<div><div>H9N2 and H5N2 avian influenza A viruses (IAVs) are two of the most widespread and economically damaging subtypes affecting the poultry industry. Controlling these viruses in poultry remains challenging, highlighting the need for a vaccine that can confer broad and effective protection across multiple avian IAV subtypes. Previously, we fused two copies of the matrix 2 ectodomain (M2e) of human and avian IAV (2xhuM2e-2xavM2e) at the C-terminus of <em>Macrobrachium rosenbergii</em> nodavirus capsid protein (NvC). This chimeric protein (NvC-2xhuM2e-2xavM2e) self-assembled into virus-like particles (VLPs), which elicited broad immune responses and activated humoral and cellular immune responses in chickens. In this study, we evaluated the protective efficacy of these chimeric VLPs in White Leghorn chickens against H9N2 and H5N2 infections. Chickens immunized with the chimeric VLPs produced a significant level of M2e antibodies that reacted with the M2e of H9N2 and H5N2 IAVs. Following viral infections, the chimeric VLPs mitigated weight loss in H9N2-infected chickens and alleviated delayed weight gain in H5N2-infected chickens. Furthermore, the chimeric VLPs reduced the clinical signs in immunized chickens challenged with either H9N2 or H5N2 avian IAV. Viral loads in the lung tissues of the immunized chickens were significantly lower compared to the negative control group. The chimeric VLPs also enhanced H9N2 and H5N2 viral clearance, showing a significant reduction in viral shedding in the oropharynges and cloacae of the immunized chickens. Furthermore, the VLPs activated both inflammatory and anti-inflammatory cytokines, which were associated with enhanced viral clearance while reducing lung and tracheal immunopathology. Collectively, these findings demonstrate the potential of the chimeric VLPs as a cross-protective vaccine against both H9N2 and H5N2 infections in poultry.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"174 ","pages":"Article 105526"},"PeriodicalIF":2.4,"publicationDate":"2025-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145562992","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}
Pub Date : 2025-11-17DOI: 10.1016/j.dci.2025.105525
Zhuojin He , Xiaomin Zheng , Lixin Ma, Zhilong Chen, Li Lin, Fei Shi
Vibrio harveyi is a major marine fish pathogen, yet the intestinal pathology and immune defense mechanisms during acute infection remain unclear. The effects of V. harveyi infection on intestinal structure, biochemical indicators, and transcriptomic profiles of hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂) were investigated. The results showed that V. harveyi infection significantly increased goblet cell hyperplasia (P < 0.05), thinning of the midgut wall (P < 0.05), and intestinal apoptosis (P < 0.05). Biochemical assays revealed significantly up-regulated activities of catalase (CAT) (P < 0.05) and superoxide dismutase (SOD) (P < 0.05), accompanied by decreased glutathione (GSH) levels (P < 0.05) and increased malondialdehyde (MDA) content (P < 0.01). Transcriptomic analysis revealed differentially expressed genes were enriched in the Antigen processing and presentation pathway and Cytokine-cytokine receptor interaction pathway. Additionally, quantitative real-time PCR demonstrated increased mRNA expression of Interleukin-6 (IL-6), Nuclear Factor Kappa-B p65 (NF-κB p65), and the tight-junction protein Claudin-3α (CLDN-3α). Redundancy analysis identified significant correlations among oxidative stress indicators, differentially expressed genes, and intestinal morphometric parameters (goblet cell number, villus length and intestinal wall thickness). Collectively, these findings highlight that V. harveyi compromises hybrid grouper health through disruption of mucosal structure, oxidative homeostasis, and immune regulation.
Pub Date : 2025-11-15DOI: 10.1016/j.dci.2025.105523
Lei Wang , Chenlong Wang , Hongyan Ding , Shibin Feng , Xichun Wang , Jinjie Wu , Dong Liu
Soybean glycinin (11S) and β-conglycinin (7S) are major contributors to allergic diarrhea and intestinal barrier damage in young animals. This study investigated the molecular mechanisms underlying the 7S- and 11S-induced dysfunction of mitochondrial and endoplasmic reticulum (ER) interactions in porcine intestinal epithelial (IPEC-J2) cells via the oxidative stress pathway. The results showed that 7S- and 11S-induced oxidative stress, as evidenced by the following findings: reduced manganese superoxide dismutase (Mn-SOD) activity and elevated 8-Hydroxy-2′-deoxyguanosine (8-OHdG) levels, with excessive reactive oxygen species (ROS) accumulation and elevated Ca2+ levels; decreased mitochondrial membrane potential (MMP), damaged mitochondria-associated endoplasmic reticulum membranes (MAM) structure; up-regulated the protein expression of glucose-regulated protein 75 (GRP75) and mitochondrial Rho-GTPase 1 (Miro1), while inositol 1,4,5 -trisphosphate receptor (IP3R), voltage-dependent anion channel 1 (VDAC1), mitofusin2 (MFN2) and phosphofurin acidic cluster sorting protein 2 (PACS2) were down-regulated. N-acetylcysteine (NAC)pre-treatment alleviated ROS accumulation and mitigated Ca2+ overload and MAM dysfunction, thereby ameliorating IPEC-J2 cell injury. In conclusion, 7S- and 11S-induced ROS burst to disrupt mitochondria-ER interaction homeostasis, leading to MAM structural damage and calcium dysregulation in IPEC-J2 cells, NAC effectively mitigated this process by scavenging ROS. These findings elucidate the critical involvement of subcellular organelle interaction disorders in food allergy pathogenesis and provide novel insights for targeted intervention strategies.
{"title":"Soybean glycinin and β-conglycinin disrupted mitochondria-endoplasmic reticulum interactions in porcine intestinal epithelial cells via oxidative stress cascades","authors":"Lei Wang , Chenlong Wang , Hongyan Ding , Shibin Feng , Xichun Wang , Jinjie Wu , Dong Liu","doi":"10.1016/j.dci.2025.105523","DOIUrl":"10.1016/j.dci.2025.105523","url":null,"abstract":"<div><div>Soybean glycinin (11S) and β-conglycinin (7S) are major contributors to allergic diarrhea and intestinal barrier damage in young animals. This study investigated the molecular mechanisms underlying the 7S- and 11S-induced dysfunction of mitochondrial and endoplasmic reticulum (ER) interactions in porcine intestinal epithelial (IPEC-J2) cells via the oxidative stress pathway. The results showed that 7S- and 11S-induced oxidative stress, as evidenced by the following findings: reduced manganese superoxide dismutase (Mn-SOD) activity and elevated 8-Hydroxy-2′-deoxyguanosine (8-OHdG) levels, with excessive reactive oxygen species (ROS) accumulation and elevated Ca<sup>2+</sup> levels; decreased mitochondrial membrane potential (MMP), damaged mitochondria-associated endoplasmic reticulum membranes (MAM) structure; up-regulated the protein expression of glucose-regulated protein 75 (GRP75) and mitochondrial Rho-GTPase 1 (Miro1), while inositol 1,4,5 -trisphosphate receptor (IP3R), voltage-dependent anion channel 1 (VDAC1), mitofusin2 (MFN2) and phosphofurin acidic cluster sorting protein 2 (PACS2) were down-regulated. N-acetylcysteine (NAC)pre-treatment alleviated ROS accumulation and mitigated Ca<sup>2+</sup> overload and MAM dysfunction, thereby ameliorating IPEC-J2 cell injury. In conclusion, 7S- and 11S-induced ROS burst to disrupt mitochondria-ER interaction homeostasis, leading to MAM structural damage and calcium dysregulation in IPEC-J2 cells, NAC effectively mitigated this process by scavenging ROS. These findings elucidate the critical involvement of subcellular organelle interaction disorders in food allergy pathogenesis and provide novel insights for targeted intervention strategies.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"173 ","pages":"Article 105523"},"PeriodicalIF":2.4,"publicationDate":"2025-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145539480","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}
Pub Date : 2025-11-15DOI: 10.1016/j.dci.2025.105524
Mahmoud M. Elnaggar , Amany Hassan , Gaber S. Abdellrazeq , Asmaa H. Mahmoud , William C. Davis
Natural killer (NK) cells are the first discovered member of innate lymphoid cells (ILCs) that play a pivotal role in host defense. They were initially identified by their ability to nonspecifically lyse certain tumor and virally infected cells. What is known about NK cells in species other than humans and mice is limited. The progress that has been made is through development and use of monoclonal antibodies (mAbs) specific to NKp46 (NCR1, CD335), a major NK cell-activating receptor that has been used as a marker to identify leukocytes with NK cell characteristics in cattle, sheep, buffaloes, pigs, horses and dogs. As described herein, the finding that the mAb that recognizes sheep and cattle NKp46 recognizes the orthologue in goats has provided an opportunity to identify a subset of leukocytes with phenotypic and functional characteristics of NK cells in goats, a profile consistent with NK cell features reported in other animal species. The study expands opportunities to conduct comparative studies on NK cells across different species and underscores the potential use of goats as an animal model in investigating NK cell biology in health and disease.
{"title":"NKp46 identifies a subset of goat leukocytes with phenotypic and functional characteristics of natural killer cells","authors":"Mahmoud M. Elnaggar , Amany Hassan , Gaber S. Abdellrazeq , Asmaa H. Mahmoud , William C. Davis","doi":"10.1016/j.dci.2025.105524","DOIUrl":"10.1016/j.dci.2025.105524","url":null,"abstract":"<div><div>Natural killer (NK) cells are the first discovered member of innate lymphoid cells (ILCs) that play a pivotal role in host defense. They were initially identified by their ability to nonspecifically lyse certain tumor and virally infected cells. What is known about NK cells in species other than humans and mice is limited. The progress that has been made is through development and use of monoclonal antibodies (mAbs) specific to NKp46 (NCR1, CD335), a major NK cell-activating receptor that has been used as a marker to identify leukocytes with NK cell characteristics in cattle, sheep, buffaloes, pigs, horses and dogs. As described herein, the finding that the mAb that recognizes sheep and cattle NKp46 recognizes the orthologue in goats has provided an opportunity to identify a subset of leukocytes with phenotypic and functional characteristics of NK cells in goats, a profile consistent with NK cell features reported in other animal species. The study expands opportunities to conduct comparative studies on NK cells across different species and underscores the potential use of goats as an animal model in investigating NK cell biology in health and disease.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"173 ","pages":"Article 105524"},"PeriodicalIF":2.4,"publicationDate":"2025-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145539425","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}
Pub Date : 2025-11-15DOI: 10.1016/j.dci.2025.105515
Ying Ye , Ce Zhang , Fengyue Guo , Siyi She , Shun Yang , Mengmeng Huang , Hui Fei
Galectins are evolutionarily conserved lectins that play pivotal roles in innate immunity. In this study, we functionally characterized a novel chimera-type galectin-3 homolog from largemouth bass (Micropterus salmoides), designated MsGal-3B. The open reading frame of MsGal-3B encodes a 258-amino acid protein containing a canonical carbohydrate recognition domain (CRD) and an N-terminal non-lectin region, typical of galectin-3 family members. Phylogenetic analysis revealed that MsGal-3B clusters with galectin-3 orthologs from Perciformes, suggesting functional conservation within this order. Recombinant MsGal-3B (rMsGal-3B) exhibited strong affinity for peptidoglycan (PGN) and β-glucan (GLU), and weak interaction with lipopolysaccharide (LPS). Microbial binding assays revealed broad recognition across Gram-positive bacteria (Micrococcus luteus, Staphylococcus aureus), Gram-negative bacteria (Aeromonas veronii, Vibrio parahaemolyticus), and fungi (Saccharomyces cerevisiae, Pichia pastoris). Notably, rMsGal-3B selectively agglutinated Escherichia coli and M. luteus, but not A. veronii or S. cerevisiae, indicating ligand-dependent functional specificity. rMsGal-3B significantly suppressed the proliferation of E. coli, M. luteus, and S. cerevisiae, while showing no inhibitory effect on A. veronii, suggesting a narrow-spectrum antimicrobial profile. Additionally, rMsGal-3B exhibited hemagglutination activity, implying a potential role in host cell recognition or immune modulation. Collectively, these findings demonstrate that MsGal-3B functions as a multifunctional immune effector in M. salmoides, capable of selective microbial recognition, agglutination, and targeted antimicrobial activity. This study provides new insights into the role of teleost galectins in innate immunity and highlights their potential as molecular targets for disease control in aquaculture.
{"title":"Functional characterization of Micropterus salmoides galectin-3 (MsGal-3B): a multifunctional immune effector with agglutination and antibacterial activities","authors":"Ying Ye , Ce Zhang , Fengyue Guo , Siyi She , Shun Yang , Mengmeng Huang , Hui Fei","doi":"10.1016/j.dci.2025.105515","DOIUrl":"10.1016/j.dci.2025.105515","url":null,"abstract":"<div><div>Galectins are evolutionarily conserved lectins that play pivotal roles in innate immunity. In this study, we functionally characterized a novel chimera-type galectin-3 homolog from largemouth bass (<em>Micropterus salmoides</em>), designated MsGal-3B. The open reading frame of MsGal-3B encodes a 258-amino acid protein containing a canonical carbohydrate recognition domain (CRD) and an N-terminal non-lectin region, typical of galectin-3 family members. Phylogenetic analysis revealed that MsGal-3B clusters with galectin-3 orthologs from Perciformes, suggesting functional conservation within this order. Recombinant MsGal-3B (rMsGal-3B) exhibited strong affinity for peptidoglycan (PGN) and β-glucan (GLU), and weak interaction with lipopolysaccharide (LPS). Microbial binding assays revealed broad recognition across Gram-positive bacteria (<em>Micrococcus luteus</em>, <em>Staphylococcus aureus</em>), Gram-negative bacteria (<em>Aeromonas veronii</em>, <em>Vibrio parahaemolyticus</em>), and fungi (<em>Saccharomyces cerevisiae</em>, <em>Pichia pastoris</em>). Notably, rMsGal-3B selectively agglutinated <em>Escherichia coli</em> and <em>M. luteus</em>, but not <em>A. veronii</em> or <em>S. cerevisiae</em>, indicating ligand-dependent functional specificity. rMsGal-3B significantly suppressed the proliferation of <em>E. coli</em>, <em>M. luteus</em>, and <em>S. cerevisiae</em>, while showing no inhibitory effect on <em>A. veronii</em>, suggesting a narrow-spectrum antimicrobial profile. Additionally, rMsGal-3B exhibited hemagglutination activity, implying a potential role in host cell recognition or immune modulation. Collectively, these findings demonstrate that MsGal-3B functions as a multifunctional immune effector in <em>M. salmoides</em>, capable of selective microbial recognition, agglutination, and targeted antimicrobial activity. This study provides new insights into the role of teleost galectins in innate immunity and highlights their potential as molecular targets for disease control in aquaculture.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"173 ","pages":"Article 105515"},"PeriodicalIF":2.4,"publicationDate":"2025-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145534362","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}
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