Developmental and comparative immunology最新文献

{"title":"Functional MDA5 knockout in CHSE-214 cells decreases the response to long dsRNA and enhances Chum Salmon Reovirus susceptibility","authors":"Shayne Oberhoffner ,&nbsp;Mathilde Peruzzi ,&nbsp;Bertrand Collet ,&nbsp;Stephanie DeWitte-Orr","doi":"10.1016/j.dci.2025.105466","DOIUrl":"10.1016/j.dci.2025.105466","url":null,"abstract":"<div><div>Many organisms express sensors that detect long dsRNA, a nucleic acid produced by almost all viruses during replication. These subcellular compartment-specific pattern recognition receptors (PRRs) detect viral replication and rapidly induce a generalist immune response mediated by type I interferon and interferon-stimulated genes (ISGs). In this study, the role of melanoma differentiation-associated protein (MDA)5, a cytoplasmic sensor of long dsRNA, was measured in CHSE-214 cells that have been genetically modified for MDA5 to either to be non-functional (frameshift mutation; MDA5C1) or have a silent point mutation (MDA5C2). These cells were transfected with long dsRNA of either 659bp or 334bp in length, and their ability to induce ISG transcript expression (<em>Mx1, vig-3</em> and <em>vig-4</em>) was measured by RT-qPCR. MDA5C1 produced significantly less <em>Mx1, vig-3</em> and <em>vig-4</em> transcripts compared to MDA5C2 when treated with the 659bp dsRNA molecule, while there was no difference between cell lines in <em>Mx1</em> expression levels when induced by a 334 bp dsRNA molecule. MDA5C1 was more susceptible to CSV than MDA5C2, and dsRNA treatment was able to protect both cell lines from CSV infection. However, there were no length dependent effects observed, nor were there differences in dsRNA-mediated protection between cell lines. This data suggests that salmonid MDA5 has a length preference similar to mammals, with longer dsRNA molecules being better ligands for this receptor. It also suggests that MDA5 may play a role in the antiviral response against CSV but the antiviral response is not dependent on MDA5. These are important findings for understanding the fundamental importance of MDA5 in the antiviral response of salmonids.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"172 ","pages":"Article 105466"},"PeriodicalIF":2.4,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145091341","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 immune responses of the noble scallop Chlamys nobilis to Vibrio parahaemolyticus infection","authors":"Jing Sun ,&nbsp;Ziru Huang ,&nbsp;Qing Lin ,&nbsp;Haixin Hu ,&nbsp;Hailong Su ,&nbsp;Taixin Chen ,&nbsp;Shitong Liu ,&nbsp;Changping Xu ,&nbsp;Yifan Li ,&nbsp;Hongkuan Zhang ,&nbsp;Huaiping Zheng","doi":"10.1016/j.dci.2025.105502","DOIUrl":"10.1016/j.dci.2025.105502","url":null,"abstract":"<div><div><em>Vibrio parahaemolyticus</em> (<em>VP</em>) represents a substantial threat to both the global aquaculture industry and public health. While, <em>VP</em> pathogenicity has been investigated in various bivalve species, the dynamic immune response of the economically important noble scallop <em>Chlamys nobilis</em> remains inadequately characterized. This study provides the first integrative analysis combining histopathological, physiological, and transcriptomic approaches to elucidate the scallop's immune response to <em>VP</em> infection across multiple time points. Our findings indicate that gill tissue damage escalates with the duration of infection. Physiological indices, including the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) levels, peaked at 24 h post-infection (<em>P</em> &lt; 0.05). Concurrently, malondialdehyde (MDA) levels exhibited a significant increase from 24 h (<em>P</em> &lt; 0.05). Transcriptomic analysis identified 5,058, 2,045, 3,952, and 6,490 differentially expressed genes (DEGs) at 6, 24, 48, and 72 h, respectively. Enrichment analyses underscored the significant regulation of pathways related to cellular structure, signal transduction (e.g., MAPK signaling pathway), and metabolism (e.g., ribosome, proteasome, and oxidative phosphorylation). Notably, key immune-related genes (e.g., <em>IRAK1</em>, <em>DDX58</em>, <em>CASP2</em>, <em>CASP7</em>, <em>XIAP</em>) were significantly down-regulated throughout the infection period, despite a transient rebound at 24 h. These results demonstrate that <em>VP</em> infection may lead to the suppression and dysregulation of the immune defenses of <em>C. nobilis</em> over time. This study advances our understanding of scallop antibacterial immunity and provides a foundation for future research on molecular immune responses in bivalves.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"172 ","pages":"Article 105502"},"PeriodicalIF":2.4,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145412767","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":"A review of host and pathogen gene targets for RNAi therapeutics in shrimp disease mitigation","authors":"Maria Violeta R. Tare ,&nbsp;Debrah Jannsen DJ N. Almazan ,&nbsp;Krisha Marie D.L. Saquilayan ,&nbsp;Patricia Jhoanna V. Glori ,&nbsp;Jim Troy Solitario ,&nbsp;Mary Beth B. Maningas","doi":"10.1016/j.dci.2025.105482","DOIUrl":"10.1016/j.dci.2025.105482","url":null,"abstract":"<div><div>RNA interference (RNAi) has emerged as a powerful therapeutic and research tool in shrimp aquaculture, with its significant potential for disease mitigation. This review explores the strategic targeting of essential genes–either from the host gene or the pathogen–as a basis for RNAi-based therapies in shrimp. Targeting host genes, especially those involved in immune responses, can lead to increased protective effects but also poses risks of lethality if essential physiological pathways are disrupted. Conversely, silencing pathogen genes offers pathogen-specific intervention, though with limitations in broad-spectrum applicability and potential off-target effects. This paper further discusses multi-variant RNAi approaches that target both host and pathogen genes, highlighting their potential in achieving greater protective effects. Also included is a review of Bioinformatics and AI applications in RNAi, and the possibilities it may offer for gene silencing in shrimp. Current evidence suggests that targeting host immune-related genes may yield higher survival outcomes, but a tailored, case-by-case approach remains important. Ultimately, the choice of target depends on a nuanced understanding of host-pathogen interactions. The findings underscore the need for continued research to optimize RNAi strategies for sustainable shrimp disease management.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"172 ","pages":"Article 105482"},"PeriodicalIF":2.4,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145211899","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":"Non-lethal heat shock enhances the immune response of Procambarus clarkii hemocytes to Vibrio parahaemolyticus through the HSP90 gene family","authors":"Shirui Yue ,&nbsp;Xiuhong Cai ,&nbsp;Zhangxuan Chen ,&nbsp;Yuyan Gong ,&nbsp;Yuqing Liu ,&nbsp;Bao Wang ,&nbsp;Yewen Xi ,&nbsp;Shunchang Wang ,&nbsp;Xin Zhang","doi":"10.1016/j.dci.2025.105484","DOIUrl":"10.1016/j.dci.2025.105484","url":null,"abstract":"<div><div>This research focuses on <em>Procambarus clarkii</em> and systematically explores the mechanism of action of the <em>HSP90</em> gene family when responding to non-lethal heat shock (NLHS) and <em>Vibrio parahaemolyticus</em> infection. We have identified four members of the <em>HSP90</em> gene family. The characteristics of the proteins encoded by these genes are significantly different, which implies functional divergence. Multiple sequence alignment and phylogenetic analysis suggested that the structure of HSP90 proteins exhibits both conservatism and diversity, indicating functional divergence among family members. The genes are unevenly distributed across chromosomes and there are no tandem repeat genes. <em>HSP90</em> genes are differentially expressed in tissues, with high expression in hemocytes indicating immune involvement. After NLHS, the expression of some genes increased dramatically and reached the peak after injection with <em>V. parahaemolyticus</em>. However, for some other genes, their expression did not change significantly, indicating that they may be involved in different types of regulation. Protein interaction and co-expression analyses revealed HSP90 proteins interact with innate immune proteins and collaborate with TLR pathway members. DsRNA silencing of <em>HSP90</em> downregulates key TLR molecules (<em>TLR2</em>, <em>TRAF6</em>, <em>IRAK4</em>, <em>MyD88</em>) at different time points, confirming their functional immune association. This indicates <em>HSP90</em> genes play a key role in anti-pathogen mechanisms. The study provides molecular markers for breeding stress-resistant <em>P. clarkii</em> and theoretical support for crustacean stress biology research.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"172 ","pages":"Article 105484"},"PeriodicalIF":2.4,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145211944","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":"Classical swine fever virus genotype 2.1 triggers stronger inflammatory and immune responses in porcine alveolar macrophages than genotype 3.4","authors":"Yu-Chun Huang ,&nbsp;Ming-Chung Deng ,&nbsp;Yu-Liang Huang ,&nbsp;Kuo-Jung Tsai ,&nbsp;Hsin-Meng Liu ,&nbsp;I-Li Liu ,&nbsp;Chen-Si Lin ,&nbsp;Chia-Yi Chang","doi":"10.1016/j.dci.2025.105496","DOIUrl":"10.1016/j.dci.2025.105496","url":null,"abstract":"<div><div>Classical swine fever (CSF), caused by classical swine fever virus (CSFV), is a highly contagious disease characterized by immunosuppression. Since 1996, a shift from CSFV genotype 3.4 (G3.4) to genotype 2.1 (G2.1) has been observed in Taiwan. Although G2.1 exhibits higher replication efficiency than G3.4, their differential immune regulatory mechanisms remain unclear. This study investigated the responses of porcine alveolar macrophages (PAMs) infected with G2.1 versus G3.4, regarding their infection rates, cytokine protein levels, and transcriptomic profiling performed by RNA-seq with qPCR validation. G2.1 showed a significantly higher infection rate at 1 day post-infection (dpi) and induced stronger pro-inflammatory responses, including elevated IL-18 (1–3 dpi) and TNF-α (2 dpi) levels. Transcriptomic analysis revealed that G2.1 enriched immune pathways, such as JAK-STAT signaling, which favor Th17 cell differentiation. In contrast, G3.4 favors IL-17 signaling and apoptotic pathways, which are associated with B-cell recruitment, potentially promoting faster viral clearance. These findings reveal that while both CSFV genotypes engage the Th17 axis, G2.1 drives upstream differentiation to establish a pro-inflammatory niche, whereas G3.4 engages terminal IL-17 responses conducive to immune resolution. This study provides new insight into genotype-specific host responses that may contribute to the CSFV genotype shift observed in the field.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"172 ","pages":"Article 105496"},"PeriodicalIF":2.4,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145336560","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":"Screening of host proteins interacting with the major capsid protein of decapod iridescent virus 1 in Litopenaeus vannamei by the yeast two-hybrid system","authors":"Yisang Lu ,&nbsp;Rongxiang Tan ,&nbsp;Jian Ye ,&nbsp;Guangjie Si ,&nbsp;Ting Xu ,&nbsp;Xiaoyi Pan","doi":"10.1016/j.dci.2025.105500","DOIUrl":"10.1016/j.dci.2025.105500","url":null,"abstract":"<div><div>Decapod iridescent virus 1 (DIV1) is a fierce pathogen causing high mortality in the decapod crustaceans, including <em>Litopenaeus vannamei.</em> Major capsid protein (MCP), the main component of the viral capsid, plays a crucial role in iridovirus infection. To gain insight into the role of MCP in DIV1 infections, a pPR3-N membrane cDNA library derived from <em>L. vannamei</em> hemocyte was established, and the bait plasmid pBT3-STE-MCP was constructed. Then, the yeast two-hybrid (Y2H) system was used to screen the host cellular proteins that interact with DIV1-MCP. Sixteen proteins were identified as the interacting proteins of MCP. Three candidate proteins, including arginine kinase, sarcoplasmic calcium-binding protein, and 40S ribosomal protein S13, were selected, and <em>in vitro</em> GST pull-down assays were performed and verified their interaction with MCP. Our findings provide valuable information for better insights into DIV1 pathogenesis and the role of MCP in this process.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"172 ","pages":"Article 105500"},"PeriodicalIF":2.4,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145367864","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":"Exploring trained immunity to complement vaccination against Streptococcus suis in swine","authors":"Lidia Sánchez-Morales ,&nbsp;Marta Pérez-Sancho ,&nbsp;Teresa García-Seco ,&nbsp;Ana Balseiro ,&nbsp;Andrea Pérez-Domingo ,&nbsp;Aránzazu Buendía ,&nbsp;Alberto Diez-Guerrier ,&nbsp;María García de Garnica García ,&nbsp;Patricia Mareque ,&nbsp;María de los Ángeles Risalde ,&nbsp;Christian Gortázar ,&nbsp;Mercedes Domínguez ,&nbsp;Lucas Domínguez","doi":"10.1016/j.dci.2025.105498","DOIUrl":"10.1016/j.dci.2025.105498","url":null,"abstract":"<div><div><em>Streptococcus suis</em> is a major pathogen in swine, causing septicaemia, meningitis, and arthritis. Effective control is complicated by the presence of multiple serotypes, genetic heterogeneity, and interference of maternal antibodies with vaccine efficacy. Moreover, the emergence of antibiotic-resistant strains poses additional challenges. Trained immunity, a process of innate immune memory induced by certain stimuli, has been proposed as a novel approach to enhance host defense mechanisms.</div><div>This study aimed to evaluate whether a mycobacterial-derived immunomodulator (dpB) administered with a specific <em>S. suis</em> vaccine could improve protection in pigs. Thirty-two weaned pigs were allocated into four groups: vaccine alone, vaccine and live dpB, vaccine and inactivated dpB, and non-immunized controls. After two immunizations, animals were challenged with a <em>S. suis</em> serotype 2 strain.</div><div>Immune responses were assessed through <em>ex vivo</em> blood stimulation, cytokine measurement and phagocytosis assays. The group receiving vaccine and inactivated dpB presented a better response, including reduced bacterial loads in tissues, less severe lung lesions and elevated pre-challenge cytokine levels. Clinically, these animals showed milder symptoms and lower bacterial dissemination post-challenge.</div><div>While trained immunity remains a relatively new concept in veterinary immunology, these results suggest that inactivated dpB, when combined with specific vaccination, could serve as an effective complementary strategy to improve protection against <em>S. suis</em>. Further research is needed to elucidate the underlying mechanisms and to establish its potential role in comprehensive disease prevention programs.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"172 ","pages":"Article 105498"},"PeriodicalIF":2.4,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145370133","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 immunomodulatory roles of a novel glucocorticoid receptor (GPR97) in the large yellow croaker (Larimichthys crocea)","authors":"Jixiu Wang ,&nbsp;Chenqian Wu ,&nbsp;Zhiqing Ye ,&nbsp;Zhijing Jiang ,&nbsp;Baoyi Huang ,&nbsp;Haojie Yuan ,&nbsp;Xudong Liang ,&nbsp;Lifei Ge ,&nbsp;Xiaolong Yin ,&nbsp;Weiye Li ,&nbsp;Xiuwen Xu ,&nbsp;Tianming Wang ,&nbsp;Jingwen Yang","doi":"10.1016/j.dci.2025.105506","DOIUrl":"10.1016/j.dci.2025.105506","url":null,"abstract":"<div><div>GPR97, a member of the adhesion G protein-coupled receptor (aGPCR) family, functions as a membrane receptor for glucocorticoids, mediating their rapid non-genomic effects and immune regulation in mammals. However, the membrane receptor for glucocorticoids in fish and the mechanisms underlying cortisol's rapid responses remain poorly understood. In this study, GPR97 was cloned and characterized from the large yellow croaker (<em>Larimichthys crocea</em>) as <em>Lc</em>GPR97. Sequence alignment and membrane localization analyses confirmed its classification within the aGPCR family. Upon cortisol stimulation in HEK293 cells expressing <em>Lc</em>GPR97, intracellular cAMP levels decreased, Ca²⁺ concentration increased, and ERK1/2 phosphorylation was activated. These results suggest that cortisol activates <em>Lc</em>GPR97, which couples with Gα_i/o proteins to mediate downstream signaling. Furthermore, <em>Lc</em>GPR97 was highly expressed in immune-related tissues, including the spleen, trunk kidney and head kidney, and its expression increased after lipopolysaccharide (LPS) stimulation, indicating its role in immune modulation during inflammation. These findings offer novel insights into the molecular mechanisms of cortisol's rapid effects via membrane receptors in fish and highlight potential implications for immune regulation in aquaculture.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"172 ","pages":"Article 105506"},"PeriodicalIF":2.4,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145387936","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":"Methyltransferase WTAP participates in shrimp defense against Vibrio parahaemolyticus infection","authors":"Yuxin Shi ,&nbsp;Jingyang Liu ,&nbsp;Tongxin Lin ,&nbsp;Wanli Lu ,&nbsp;Linwei Yang ,&nbsp;Siyou Huang","doi":"10.1016/j.dci.2025.105510","DOIUrl":"10.1016/j.dci.2025.105510","url":null,"abstract":"<div><div>Although the immune function of the methyltransferase Wilm's tumor 1-associated protein (WTAP) has been well-studied in vertebrates, its role in the invertebrate immune system remains limited. In this study, we identified the <em>WTAP</em> gene in <em>Penaeus vannamei</em> and investigated its potential role in shrimp innate immunity. RT-qPCR analysis revealed that <em>WTAP</em> was broadly expressed in key immune tissues of the shrimp, including hemocytes, intestines, hepatopancreas, and gills. <em>WTAP</em> expression was upregulated in the hepatopancreas and gills during <em>Vibrio parahaemolyticus</em> infection. Although knockdown of <em>WTAP</em> did not significantly affect the survival rate of shrimp infected with <em>V. parahaemolyticus</em>, there was a non-significant trend toward decreased survival, along with markedly increased bacterial loads in gills and hepatopancreas. Knockdown of <em>WTAP</em> also led to a significant reduction in the expression levels of <em>ALF1/3/5/6</em> and <em>PEN2/3/4</em>. Furthermore, <em>WTAP</em> expression was regulated by NF-κB Relish and repressed by the transcription factors STAT and Dorsal. Our results demonstrated that <em>P. vannamei</em> WTAP, regulated by NF-κB Relish, inhibited <em>V. parahaemolyticus</em> proliferation by modulating the expression of antimicrobial peptides (AMPs). These findings deepen our understanding of WTAP's role in crustacean innate immunity and provide a crucial foundation for future research on the relationship between m⁶A modification and immune regulation in invertebrates.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"172 ","pages":"Article 105510"},"PeriodicalIF":2.4,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145457732","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":"Screening for C-type lectin receptor (CLR)/bacteria interactions using a bovine CLR-Fc fusion protein library reveals recognition of Pasteurella multocida B:2 by MICL","authors":"Samira Christin Görig ,&nbsp;Yeliz Gün ,&nbsp;Dimitri Leonid Lindenwald ,&nbsp;Jochen Meens ,&nbsp;Hans-Joachim Schuberth ,&nbsp;Bernd Lepenies","doi":"10.1016/j.dci.2025.105474","DOIUrl":"10.1016/j.dci.2025.105474","url":null,"abstract":"<div><div>Pattern recognition receptors (PRRs) are an essential component of the innate immune system. Myeloid C-type-lectin receptors (CLRs) serve as PRRs and play a crucial role in pathogen recognition. While the role of CLRs has been mainly studied in mice and humans, their function in cattle is poorly understood. To address this gap, we generated a novel bovine CLR-hFc fusion protein library, enabling high-throughput screening of bovine CLR/pathogen interactions.</div><div>The functionality of the bovine CLR-hFc fusion proteins was validated with known CLR ligands using ELISA- and flow cytometry-based binding assays, by comparison of bovine CLRs with their murine, ovine and human orthologues. In a proof-of-principle pathogen binding study, we assessed CLR binding to <em>Pasteurella (P.) multocida,</em> a Gram-negative bacterial pathogen causing hemorrhagic septicemia in cattle. The bovine CLR myeloid inhibitory C-type lectin (MICL, Clec12A) was identified as a potential receptor for <em>P. multocida</em>, as it exhibited significant binding in flow cytometry binding assays. Cross-species analysis confirmed that murine and ovine MICL also binds <em>P. multocida</em>, suggesting an evolutionarily conserved recognition.</div><div>To explore MICL-dependent innate responses to <em>P. multocida</em>-derived factors, cytokine assays were performed using dendritic cells (DCs) from wild-type (WT) and MICL-deficient (MICL^−/−) mice. MICL^−/− DCs produced higher levels of IL-6 and IL-12 upon stimulation with heat-killed <em>P. multocida</em>, suggesting a role for MICL in the down-modulation of innate responses.</div><div>The results highlight MICL as a receptor in the recognition of <em>P. multocida</em> and demonstrate the utility of the generated bovine CLR-hFc fusion protein library for pathogen screening.</div></div>","PeriodicalId":11228,"journal":{"name":"Developmental and comparative immunology","volume":"172 ","pages":"Article 105474"},"PeriodicalIF":2.4,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145124514","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}