The neonatal Fc receptor (FcRn) binds to IgG CH2 and CH3 domains (the Fc segment), triggering transendocytosis. Therefore, FcRn transports biological agents across the mucosal barrier. Mucosal administration provides less stimulation to the body than other methods. However, whether FcRn is an effective carrier for antigens across bovine respiratory epithelial cells is unknown. Here, an antigen was fused with the Fc fragment and transferred through the mucosal barrier to antigen-presenting cells via active transport mediated by FcRn. We established a model of FcRn-mediated recombinant IgG Fc protein expression in bovine embryonic tracheal epithelial cells. Western blotting showed that SPA inhibited the relative transport amount of FcRn-mediated IgG Fc fusion protein. Fc fusion protein positively correlated with protein concentration and action time, with the maximum level reached at 1.4 mg/mL (protein concentration) and 18 h (action time). An FcRn-mediated transport model of the IgG Fc recombinant protein in guinea pig lungs was established, and the amount of protein transported at different time points was measured using immunohistochemistry. FcRn mediates vaccine antigen delivery through the mucosal barrier to activate immune cells in the lamina propria, laying a theoretical foundation for the clinical application of nasal mucosal immune vaccines.
{"title":"Neonatal Fc receptor participates in endocytosis of Fc fusion protein in vivo and in vitro","authors":"Yaping Zhou , Yanfang Wang , Hongmei Zhao , Ting Guo , Yongqing Hao","doi":"10.1016/j.vetimm.2025.110930","DOIUrl":"10.1016/j.vetimm.2025.110930","url":null,"abstract":"<div><div>The neonatal Fc receptor (FcRn) binds to IgG CH2 and CH3 domains (the Fc segment), triggering transendocytosis. Therefore, FcRn transports biological agents across the mucosal barrier. Mucosal administration provides less stimulation to the body than other methods. However, whether FcRn is an effective carrier for antigens across bovine respiratory epithelial cells is unknown. Here, an antigen was fused with the Fc fragment and transferred through the mucosal barrier to antigen-presenting cells via active transport mediated by FcRn. We established a model of FcRn-mediated recombinant IgG Fc protein expression in bovine embryonic tracheal epithelial cells. Western blotting showed that SPA inhibited the relative transport amount of FcRn-mediated IgG Fc fusion protein. Fc fusion protein positively correlated with protein concentration and action time, with the maximum level reached at 1.4 mg/mL (protein concentration) and 18 h (action time). An FcRn-mediated transport model of the IgG Fc recombinant protein in guinea pig lungs was established, and the amount of protein transported at different time points was measured using immunohistochemistry. FcRn mediates vaccine antigen delivery through the mucosal barrier to activate immune cells in the lamina propria, laying a theoretical foundation for the clinical application of nasal mucosal immune vaccines.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"283 ","pages":"Article 110930"},"PeriodicalIF":1.4,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143777551","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}
Pub Date : 2025-05-01Epub Date: 2025-04-03DOI: 10.1016/j.vetimm.2025.110934
Pham Hoang Son Hung, Ho Thi Dung, Le Duc Thao, Nguyen Van Chao, Nguyen Thi Hoa, Bui Thi Hien, Anjan Mondal, Victor Nsereko, Le Dinh Phung
This study evaluates the effects of Saccharomyces cerevisiae fermentation-derived postbiotics (SCFP) supplementation on diarrhea incidence, small intestinal morphology, and expression of tight junction genes in piglets. The study compared three groups: a control group (CON), which received a standard basal diet; a standard basal control diet containing 1.0 kg/mT of Beta-glucan 50 % (BG); and a standard basal control diet containing 2.0 kg/mT of SCFP (Diamond V XPC). The experimental design involved feeding the diets to the sows from the day when they were inseminated until their piglets were weaned and to piglets from birth to weaning. Diarrhea incidence was monitored, intestinal morphology was assessed, and gene expression of tight junction proteins (Claudin-1, Claudin-2, Occludin, and ZO-1) and inflammatory cytokines (IL-1β) was analyzed using qPCR. Results revealed that SCFP supplementation significantly reduced diarrhea incidence and upregulated the expression of tight junction proteins Claudin-1 (1.61-fold) and Occludin (1.90-fold) compared to CON. These improvements were not associated with changes in intestinal morphology. BG supplementation showed intermediate effects on tight junction gene expression but did not differ significantly from CON. These findings highlight the potential of SCFP as a dietary supplement to enhance gastrointestinal health in piglets by strengthening the intestinal epithelial barrier and reducing pathogen translocation. The study underscores the efficacy of SCFP in improving gut health without altering intestinal structure, offering an effective approach to manage pre-weaning diarrhea. Future studies are needed to explore the long-term impact of SCFP on growth performance and immunity.
本研究评价了添加酿酒酵母发酵后生物制剂（SCFP）对仔猪腹泻发生率、小肠形态和紧密连接基因表达的影响。该研究比较了三组：对照组（CON），接受标准的基础饮食；标准基础对照日粮，含有1.0 kg/mT的β -葡聚糖50% % (BG)；标准基础对照饲粮中添加2.0 kg/mT的SCFP （Diamond V XPC）。试验设计包括从母猪授精之日起至仔猪断奶，以及仔猪从出生到断奶。监测腹泻发生率，评估肠道形态，并采用qPCR分析紧密连接蛋白（Claudin-1、Claudin-2、Occludin和ZO-1）和炎症因子（IL-1β）的基因表达。结果显示，与对照组相比，添加SCFP显著降低了腹泻发生率，上调了紧密连接蛋白Claudin-1（1.61倍）和Occludin（1.90倍）的表达，这些改善与肠道形态的变化无关。添加BG对紧密连接基因表达有中间影响，但与con没有显著差异。这些发现表明，SCFP作为一种饲粮添加剂，有可能通过增强肠上皮屏障和减少病原体易位来改善仔猪的胃肠道健康。该研究强调了SCFP在不改变肠道结构的情况下改善肠道健康的功效，为治疗断奶前腹泻提供了一种有效的方法。需要进一步的研究来探索SCFP对生长性能和免疫的长期影响。
{"title":"Effects of Saccharomyces cerevisiae fermentation-derived postbiotics supplementation in sows and piglets' diet on intestinal morphology, and intestinal barrier function in weaned pigs in an intensive pig production system","authors":"Pham Hoang Son Hung, Ho Thi Dung, Le Duc Thao, Nguyen Van Chao, Nguyen Thi Hoa, Bui Thi Hien, Anjan Mondal, Victor Nsereko, Le Dinh Phung","doi":"10.1016/j.vetimm.2025.110934","DOIUrl":"10.1016/j.vetimm.2025.110934","url":null,"abstract":"<div><div>This study evaluates the effects of <em>Saccharomyces cerevisiae</em> fermentation-derived postbiotics (SCFP) supplementation on diarrhea incidence, small intestinal morphology, and expression of tight junction genes in piglets. The study compared three groups: a control group (CON), which received a standard basal diet; a standard basal control diet containing 1.0 kg/mT of Beta-glucan 50 % (BG); and a standard basal control diet containing 2.0 kg/mT of SCFP (Diamond V XPC). The experimental design involved feeding the diets to the sows from the day when they were inseminated until their piglets were weaned and to piglets from birth to weaning. Diarrhea incidence was monitored, intestinal morphology was assessed, and gene expression of tight junction proteins (Claudin-1, Claudin-2, Occludin, and ZO-1) and inflammatory cytokines (IL-1β) was analyzed using qPCR. Results revealed that SCFP supplementation significantly reduced diarrhea incidence and upregulated the expression of tight junction proteins Claudin-1 (1.61-fold) and Occludin (1.90-fold) compared to CON. These improvements were not associated with changes in intestinal morphology. BG supplementation showed intermediate effects on tight junction gene expression but did not differ significantly from CON. These findings highlight the potential of SCFP as a dietary supplement to enhance gastrointestinal health in piglets by strengthening the intestinal epithelial barrier and reducing pathogen translocation. The study underscores the efficacy of SCFP in improving gut health without altering intestinal structure, offering an effective approach to manage pre-weaning diarrhea. Future studies are needed to explore the long-term impact of SCFP on growth performance and immunity.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"283 ","pages":"Article 110934"},"PeriodicalIF":1.4,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143768565","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 : 2025-05-01Epub Date: 2025-03-27DOI: 10.1016/j.vetimm.2025.110922
Shaojue Guo , Zhao Shuaiying , Kong Yingying , Junming Tang , Jianfeng Xu , Yuanyuan Dai , Yong Geng
Objective
RSPO2 (R-spondin 2) is a key regulator of the Wnt/β-catenin signaling pathway, involved in embryogenesis, tissue homeostasis, and cancer progression. Despite its therapeutic potential, effective agents targeting RSPO2 remain elusive. To address the unmet need for RSPO2-targeted therapies, we aimed to develop high-affinity nanobodies via phage display and prokaryotic expression, characterizing their binding specificity and functional blockade of RSPO2-LGR4 interactions. This study provides foundational insights into nanobody-mediated inhibition of Wnt signaling, supporting future therapeutic strategies against RSPO2-driven pathologies.
Methods
Recombinant RSPO2 proteins were constructed and purified using PCR-based recombination. Camels (Camelus bactrianus) were immunized with RSPO2, and phage display was employed to screen nanobody libraries. High-affinity nanobodies were cloned, expressed, purified, and assessed for specificity and binding affinity using biolayer interferometry and protein blotting. Functional validation was performed using TOPFLASH assays to evaluate their impact on Wnt/β-catenin signaling.
Results
Nanobodies with high specificity and nanomolar-range affinity constants (KDs) for RSPO2 were identified. The nanobody effectively inhibited RSPO2-induced Wnt/β-catenin signaling in human renal epithelial cells.
Conclusion
The development of RSPO2-targeting nanobodies offers new prospects for treating RSPO2-related diseases. The nanobody serve as valuable tools for functional research and hold potential as diagnostic and therapeutic agents for RSPO2-driven conditions.
{"title":"Screening, expression, and functional validation of camelid-derived nanobodies targeting RSPO2","authors":"Shaojue Guo , Zhao Shuaiying , Kong Yingying , Junming Tang , Jianfeng Xu , Yuanyuan Dai , Yong Geng","doi":"10.1016/j.vetimm.2025.110922","DOIUrl":"10.1016/j.vetimm.2025.110922","url":null,"abstract":"<div><h3>Objective</h3><div>RSPO2 (<em>R-spondin 2</em>) is a key regulator of the Wnt/β-catenin signaling pathway, involved in embryogenesis, tissue homeostasis, and cancer progression. Despite its therapeutic potential, effective agents targeting RSPO2 remain elusive. To address the unmet need for RSPO2-targeted therapies, we aimed to develop high-affinity nanobodies via phage display and prokaryotic expression, characterizing their binding specificity and functional blockade of RSPO2-LGR4 interactions. This study provides foundational insights into nanobody-mediated inhibition of Wnt signaling, supporting future therapeutic strategies against RSPO2-driven pathologies.</div></div><div><h3>Methods</h3><div>Recombinant RSPO2 proteins were constructed and purified using PCR-based recombination. Camels (<em>Camelus bactrianus</em>) were immunized with RSPO2, and phage display was employed to screen nanobody libraries. High-affinity nanobodies were cloned, expressed, purified, and assessed for specificity and binding affinity using biolayer interferometry and protein blotting. Functional validation was performed using TOPFLASH assays to evaluate their impact on Wnt/β-catenin signaling.</div></div><div><h3>Results</h3><div>Nanobodies with high specificity and nanomolar-range affinity constants (KDs) for RSPO2 were identified. The nanobody effectively inhibited RSPO2-induced Wnt/β-catenin signaling in human renal epithelial cells.</div></div><div><h3>Conclusion</h3><div>The development of RSPO2-targeting nanobodies offers new prospects for treating RSPO2-related diseases. The nanobody serve as valuable tools for functional research and hold potential as diagnostic and therapeutic agents for RSPO2-driven conditions.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"283 ","pages":"Article 110922"},"PeriodicalIF":1.4,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143759942","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 : 2025-05-01Epub Date: 2025-03-08DOI: 10.1016/j.vetimm.2025.110919
Maysa Serpa Gonçalves , Marina Martins de Oliveira , Eduarda Moraes Magossi Silva , Lorena Batalha de Souza , Rafaella Silva Andrade , Dircéia Aparecida da Costa Custódio , Amanda Carvalho Rosado Ferreira , Anna Cecília Trolesi Reis Borges Costa , Helbert Resende Freire , Carine Rodrigues Pereira , Izabela Regina Cardoso de Oliveira , Júlio Silvio de Sousa Bueno Filho , Andrey Pereira Lage , Elaine Maria Seles Dorneles
Vaccination of bovine calves is one of the main policies for bovine brucellosis control in endemic areas. However, the effect of animal age on vaccine immunogenicity is still unknown and could help to determine an ideal age for vaccination, in order to maximize immune response. Thus, the objective of this study was to compare the in vitro expression of IFN-γ by stimulated PBMC after vaccination with B. abortus S19 and RB51 strains in calves vaccinated at different ages, between 3 and 8 months. Cell-mediated immune response was assessed through culture of peripheral blood mononuclear cells (PBMC) and quantification of IFN-γ in the supernatant by enzyme-linked immunosorbent assay (ELISA). In addition, serological assays were performed using 2-mercaptoethanol (2-ME), Standard Tube Agglutination (STAT) and Fluorescent polarization assay (FPA) tests. Blood samples and sera were collected in the inoculation day, as well as at 28 and 56 days after vaccination. A generalized linear mixed model was used to evaluate effect of age at vaccination on in vitro production of IFN-γ and no differences were observed comparing the different ages, for both RB51 and S19 vaccines (p > 0.05). A higher percentage of animals vaccinated with S19 at 3–4 months-old [77.28 % (7/9)] returned to the serological negative status at day 56, when compared to 5–6-months [50 % (5/10)] and 7–8 months-old animals [27.28 % (3/11)]. In conclusion, our findings indicated similar levels of IFN-γ in vitro production in animals between 3 and 8 months of age, following vaccination with S19 and RB51 strains.
{"title":"Short communication: Effects of age on the immune response induced by Brucella abortus S19 or RB51 vaccination in calves","authors":"Maysa Serpa Gonçalves , Marina Martins de Oliveira , Eduarda Moraes Magossi Silva , Lorena Batalha de Souza , Rafaella Silva Andrade , Dircéia Aparecida da Costa Custódio , Amanda Carvalho Rosado Ferreira , Anna Cecília Trolesi Reis Borges Costa , Helbert Resende Freire , Carine Rodrigues Pereira , Izabela Regina Cardoso de Oliveira , Júlio Silvio de Sousa Bueno Filho , Andrey Pereira Lage , Elaine Maria Seles Dorneles","doi":"10.1016/j.vetimm.2025.110919","DOIUrl":"10.1016/j.vetimm.2025.110919","url":null,"abstract":"<div><div>Vaccination of bovine calves is one of the main policies for bovine brucellosis control in endemic areas. However, the effect of animal age on vaccine immunogenicity is still unknown and could help to determine an ideal age for vaccination, in order to maximize immune response. Thus, the objective of this study was to compare the <em>in vitro</em> expression of IFN-γ by stimulated PBMC after vaccination with <em>B. abortus</em> S19 and RB51 strains in calves vaccinated at different ages, between 3 and 8 months. Cell-mediated immune response was assessed through culture of peripheral blood mononuclear cells (PBMC) and quantification of IFN-γ in the supernatant by enzyme-linked immunosorbent assay (ELISA). In addition, serological assays were performed using 2-mercaptoethanol (2-ME), Standard Tube Agglutination (STAT) and Fluorescent polarization assay (FPA) tests. Blood samples and sera were collected in the inoculation day, as well as at 28 and 56 days after vaccination. A generalized linear mixed model was used to evaluate effect of age at vaccination on <em>in vitro</em> production of IFN-γ and no differences were observed comparing the different ages, for both RB51 and S19 vaccines (p > 0.05). A higher percentage of animals vaccinated with S19 at 3–4 months-old [77.28 % (7/9)] returned to the serological negative status at day 56, when compared to 5–6-months [50 % (5/10)] and 7–8 months-old animals [27.28 % (3/11)]. In conclusion, our findings indicated similar levels of IFN-γ <em>in vitro</em> production in animals between 3 and 8 months of age, following vaccination with S19 and RB51 strains.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"283 ","pages":"Article 110919"},"PeriodicalIF":1.4,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143637142","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}
IgY technology refers to the process of producing, extracting, and utilizing IgY antibodies from the egg yolk. The extraction of IgY from the avian egg yolk is particularly interesting due to its potential as a reservoir of targeted antibodies for infection prevention. The objective of our study was to evaluate and compare the efficiency and purity of antibodies obtained through four different IgY extraction procedures: water dilution (WD), polyethylene glycol (PEG) precipitation, chloroform (CF) extraction combined with PEG extraction, and phenol (PHE) extraction. The WD and CF techniques exhibited increased protein quantities; however, the reported IgY purity was reduced due to impurities detected in the SDS-PAGE analysis. The PEG technique provided a well-balanced approach with moderate protein content and greater purity than the WD and CF approaches. The phenol extraction procedure resulted in the best level of purity for IgY; however, the yield was lower. The WD methods and PEG precipitation extraction methods offer an effective and practical approach to purifying IgY antibodies. It is suitable for efficiently producing high-quality IgY on a wide scale following purification. The functionality of the IgY molecule is unaffected by various extraction techniques throughout an ELISA demonstration. This comparative study aims to gather valuable observational data on various IgY extraction methods. The main aim is to optimize these approaches to effectively address the specific demands of different practical working situations and antibody applications.
{"title":"Comprehensive analysis of the major IgY antibody extraction strategies from chicken egg yolk","authors":"Dhruvi Patel , Gireesh Babu K. , Sreenivasa Nayaka , Amel Gacem , Pankaj Kumar , Apurva Sharma , Krishna Kumar Yadav , Lamjed Mansour , Haresh S. Kalasariya","doi":"10.1016/j.vetimm.2025.110928","DOIUrl":"10.1016/j.vetimm.2025.110928","url":null,"abstract":"<div><div>IgY technology refers to the process of producing, extracting, and utilizing IgY antibodies from the egg yolk. The extraction of IgY from the avian egg yolk is particularly interesting due to its potential as a reservoir of targeted antibodies for infection prevention. The objective of our study was to evaluate and compare the efficiency and purity of antibodies obtained through four different IgY extraction procedures: water dilution (WD), polyethylene glycol (PEG) precipitation, chloroform (CF) extraction combined with PEG extraction, and phenol (PHE) extraction. The WD and CF techniques exhibited increased protein quantities; however, the reported IgY purity was reduced due to impurities detected in the SDS-PAGE analysis. The PEG technique provided a well-balanced approach with moderate protein content and greater purity than the WD and CF approaches. The phenol extraction procedure resulted in the best level of purity for IgY; however, the yield was lower. The WD methods and PEG precipitation extraction methods offer an effective and practical approach to purifying IgY antibodies. It is suitable for efficiently producing high-quality IgY on a wide scale following purification. The functionality of the IgY molecule is unaffected by various extraction techniques throughout an ELISA demonstration. This comparative study aims to gather valuable observational data on various IgY extraction methods. The main aim is to optimize these approaches to effectively address the specific demands of different practical working situations and antibody applications.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"283 ","pages":"Article 110928"},"PeriodicalIF":1.4,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143777552","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 : 2025-05-01Epub Date: 2025-03-27DOI: 10.1016/j.vetimm.2025.110923
Y.J. Lim , M.S. Lim , J.J. Lee , H. Bae , Y.J. Baek , G.S. Kim , Y. An , S.K. Kim , D. Yu
Interleukin-15 (IL-15) is a pleiotropic cytokine that plays a pivotal role in innate and adaptive immunity. Therefore, it is a promising therapeutic agent for cancer treatment. Despite growing interest in the use of IL-15 as an immunotherapeutic agent, there have been very few reports on its immunological and clinical effects in canine cancers. In this study, we generated recombinant canine IL-15 (rcIL-15) and evaluated its clinical and immunomodulatory effects in combination with metronomic cyclophosphamide in 15 canines with various tumor types. The treatment outcomes were assessed in a prospective clinical trial. Low-dose cyclophosphamide (12.5 mg/m2, PO, SID) was continuously administered for 8 weeks. Starting on day 14, after administering cyclophosphamide, rcIL-15 (20 μg/kg daily) was injected intravenously for 8 days. The disease control rate for combination therapy was 66.6 %, with the most notable partial response accounting for 33.3 % of hematological malignancies. The adverse events were minimal and primarily of grade 1 severity. Moreover, rcIL-15 administration led to significant elevations in anticancer lymphocyte subsets, such as natural killer and cytotoxic T cells, along with increased Ki-67 expression, indicating cellular proliferation. These changes were correlated with improved clinical outcomes. Our findings underscore the therapeutic potential and safety of combining rcIL-15 and metronomic cyclophosphamide for the treatment of various canine cancers.
{"title":"Evaluation of clinical and immunological responses to recombinant canine interleukin-15 therapy in dogs with cancer: A pilot study","authors":"Y.J. Lim , M.S. Lim , J.J. Lee , H. Bae , Y.J. Baek , G.S. Kim , Y. An , S.K. Kim , D. Yu","doi":"10.1016/j.vetimm.2025.110923","DOIUrl":"10.1016/j.vetimm.2025.110923","url":null,"abstract":"<div><div>Interleukin-15 (IL-15) is a pleiotropic cytokine that plays a pivotal role in innate and adaptive immunity. Therefore, it is a promising therapeutic agent for cancer treatment. Despite growing interest in the use of IL-15 as an immunotherapeutic agent, there have been very few reports on its immunological and clinical effects in canine cancers. In this study, we generated recombinant canine IL-15 (rcIL-15) and evaluated its clinical and immunomodulatory effects in combination with metronomic cyclophosphamide in 15 canines with various tumor types. The treatment outcomes were assessed in a prospective clinical trial. Low-dose cyclophosphamide (12.5 mg/m<sup>2</sup>, PO, SID) was continuously administered for 8 weeks. Starting on day 14, after administering cyclophosphamide, rcIL-15 (20 μg/kg daily) was injected intravenously for 8 days. The disease control rate for combination therapy was 66.6 %, with the most notable partial response accounting for 33.3 % of hematological malignancies. The adverse events were minimal and primarily of grade 1 severity. Moreover, rcIL-15 administration led to significant elevations in anticancer lymphocyte subsets, such as natural killer and cytotoxic T cells, along with increased Ki-67 expression, indicating cellular proliferation. These changes were correlated with improved clinical outcomes. Our findings underscore the therapeutic potential and safety of combining rcIL-15 and metronomic cyclophosphamide for the treatment of various canine cancers.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"283 ","pages":"Article 110923"},"PeriodicalIF":1.4,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143790949","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}
Leptospirosis is a major public health problem in humans and animals worldwide. The variable carboxy-terminal domain 7–13 of LigA (LigAc) is currently the most promising immunogen for the leptospirosis subunit vaccine. Its protective evidence was investigated in susceptible hamsters whose immunity was mostly based on the knowledge of resistant mice. The difference in immunity of these two animals might be an obstacle to successful vaccine development. The protective immunity induced by LigAc was reported to be specific antibodies while T-cell-mediated immunity has never been investigated. We reported for the first time that hamsters and mice gave dissimilar T-cell responses. Mice and hamsters were divided into 3 groups: an adjuvant plus recombinant LigAc (rLigAc) immunized, an adjuvant-injected, and a negative control group. Immunizations were done three times at 2-week intervals. The rLigAc-specific IgG antibody titers in rLigAc immunized mice and hamsters were significantly higher than in the control groups but no significant difference between the animals. The percentages of hamster CD4+ T cells were significantly higher than those of mice. Mouse CD25+CD4+ T cells responded to rLigAc significantly higher than hamsters. Interestingly, the rLigAc significantly reduced the percentage of IFN-γ+CD4+ cells in mice (≅30 %) and more decrease (≅70 %) was found in hamsters. Remarkably, it also reduced considerably hamster IL-4+CD4+ T cells (≅80 %) but an extremely low decrease in mice (≅20 %). Our result indicated that mice and hamsters gave different responses to leptospiral antigens which might be the possible key that plays a role in the outcome of disease.
{"title":"Hamster and mouse CD25+CD4+ T cell responses to the C-terminal of leptospiral Ig-like protein A","authors":"Jittima Duangsri , Chotima Potisap , Teerasit Techawiwattanaboon , Kanitha Patarakul , Rasana W. Sermswan , Surasakdi Wongratanacheewin","doi":"10.1016/j.vetimm.2025.110920","DOIUrl":"10.1016/j.vetimm.2025.110920","url":null,"abstract":"<div><div>Leptospirosis is a major public health problem in humans and animals worldwide. The variable carboxy-terminal domain 7–13 of LigA (LigAc) is currently the most promising immunogen for the leptospirosis subunit vaccine. Its protective evidence was investigated in susceptible hamsters whose immunity was mostly based on the knowledge of resistant mice. The difference in immunity of these two animals might be an obstacle to successful vaccine development. The protective immunity induced by LigAc was reported to be specific antibodies while T-cell-mediated immunity has never been investigated. We reported for the first time that hamsters and mice gave dissimilar T-cell responses. Mice and hamsters were divided into 3 groups: an adjuvant plus recombinant LigAc (rLigAc) immunized, an adjuvant-injected, and a negative control group. Immunizations were done three times at 2-week intervals. The rLigAc-specific IgG antibody titers in rLigAc immunized mice and hamsters were significantly higher than in the control groups but no significant difference between the animals. The percentages of hamster CD4<sup>+</sup> T cells were significantly higher than those of mice. Mouse CD25<sup>+</sup>CD4<sup>+</sup> T cells responded to rLigAc significantly higher than hamsters. Interestingly, the rLigAc significantly reduced the percentage of IFN-<sup>γ+</sup>CD4<sup>+</sup> cells in mice (≅30 %) and more decrease (≅70 %) was found in hamsters. Remarkably, it also reduced considerably hamster IL<sup>-</sup>4<sup>+</sup>CD4<sup>+</sup> T cells (≅80 %) but an extremely low decrease in mice (≅20 %). Our result indicated that mice and hamsters gave different responses to leptospiral antigens which might be the possible key that plays a role in the outcome of disease.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"283 ","pages":"Article 110920"},"PeriodicalIF":1.4,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143684187","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}
We report on the establishment of the unique canine multiple myeloma cloned cell lines B4–100 and B4-C4, established from the peripheral blood of a canine patient with suspected lymphoma. The cloned cells were analyzed for morphologic traits, proliferation rates, cell doubling times, as well as canine immunoglobulin production by flow cytometry. The cells were found to express IgE in the cell lysate by western blotting but did not express HGPRT, and were unable to grow in hypoxanthine-aminopterin-thymidine medium. When the cells were fused with canine peripheral blood mononuclear cells in vitro, hybridomas producing canine immunoglobulins in the culture supernatant could be generated. To our knowledge, this is the first report on establishment of canine myeloma cell lines and we submit that these cell lines may provide opportunities for the production of fully caninized antibodies with potential for therapeutic applications.
{"title":"Establishment of B4-100 and B4-C4, clonal canine multiple myeloma cell lines and their application in generating monoclonal antibody-producing fully canine hybridomas","authors":"Mark J. Micallef , Kaori Torihama , Kohei Fujikake , Akiko Kumagai , Atsushi Tanaka , Tomoyuki Kawai , Kazunori Ueda , Gakuto Hayashi , Shoji Ogino , Toshihiro Tsukui , Kenichi Masuda","doi":"10.1016/j.vetimm.2025.110925","DOIUrl":"10.1016/j.vetimm.2025.110925","url":null,"abstract":"<div><div>We report on the establishment of the unique canine multiple myeloma cloned cell lines B4–100 and B4-C4, established from the peripheral blood of a canine patient with suspected lymphoma. The cloned cells were analyzed for morphologic traits, proliferation rates, cell doubling times, as well as canine immunoglobulin production by flow cytometry. The cells were found to express IgE in the cell lysate by western blotting but did not express HGPRT, and were unable to grow in hypoxanthine-aminopterin-thymidine medium. When the cells were fused with canine peripheral blood mononuclear cells <em>in vitro</em>, hybridomas producing canine immunoglobulins in the culture supernatant could be generated. To our knowledge, this is the first report on establishment of canine myeloma cell lines and we submit that these cell lines may provide opportunities for the production of fully caninized antibodies with potential for therapeutic applications.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"283 ","pages":"Article 110925"},"PeriodicalIF":1.4,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143738798","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}
In dairy cattle, genetic selection for higher antibody-mediated (AMIR) and cell-mediated (CMIR) immune responses can enhance disease resistance. Cattle produce a unique subset of B cells with B cell receptors with ultralong complementarity determining regions 3 (CDR3). Antibodies with these specialized structures have superior virus neutralization characteristics. Published studies of B cell receptors with ultralong CDR3s in dairy cattle have been limited by the number of animals examined (1–4 animals in each study), and by varying breeds and ages. The objective of this study was to assess the percentage of IgM and IgG sequences with ultralong CDR3s, and gene usage in blood and colostral lymphocytes from cows classified as high, average, and low immune responders based on their estimated breeding values. B lymphocytes were isolated from the blood of 14 heifers and 7 cows. In addition, cells were isolated from colostrum of the 7 cows. RNA was extracted, cDNA was produced, and IgM and IgG transcripts were amplified using polymerase chain reactions. Amplicons were sequenced using Oxford Nanopore long-read sequencing. In sequences derived from blood B cells, AMIR estimated breeding values were significantly and positively associated with higher percentages of IgG ultralong CDR3 sequences. High AMIR cows (n = 3) also produced colostrum with a significantly greater percentage of IgG ultralong CDR3 sequences (18.0 %) than average AMIR cows (n = 4, mean 8.8 %). Larger studies are needed to investigate the association between percentages of B cells expressing IgG ultralong CDR3s and observed health traits.
{"title":"Blood and colostral IgM and IgG B cell repertoires in high, average, and low immune responder Holstein Friesian cows and heifers","authors":"T.E. Altvater-Hughes , H.P. Hodgins , D.C. Hodgins , C.A. Bauman , B.A. Mallard","doi":"10.1016/j.vetimm.2025.110926","DOIUrl":"10.1016/j.vetimm.2025.110926","url":null,"abstract":"<div><div>In dairy cattle, genetic selection for higher antibody-mediated (AMIR) and cell-mediated (CMIR) immune responses can enhance disease resistance. Cattle produce a unique subset of B cells with B cell receptors with ultralong complementarity determining regions 3 (CDR3). Antibodies with these specialized structures have superior virus neutralization characteristics. Published studies of B cell receptors with ultralong CDR3s in dairy cattle have been limited by the number of animals examined (1–4 animals in each study), and by varying breeds and ages. The objective of this study was to assess the percentage of IgM and IgG sequences with ultralong CDR3s, and gene usage in blood and colostral lymphocytes from cows classified as high, average, and low immune responders based on their estimated breeding values. B lymphocytes were isolated from the blood of 14 heifers and 7 cows. In addition, cells were isolated from colostrum of the 7 cows. RNA was extracted, cDNA was produced, and IgM and IgG transcripts were amplified using polymerase chain reactions. Amplicons were sequenced using Oxford Nanopore long-read sequencing. In sequences derived from blood B cells, AMIR estimated breeding values were significantly and positively associated with higher percentages of IgG ultralong CDR3 sequences. High AMIR cows (n = 3) also produced colostrum with a significantly greater percentage of IgG ultralong CDR3 sequences (18.0 %) than average AMIR cows (n = 4, mean 8.8 %). Larger studies are needed to investigate the association between percentages of B cells expressing IgG ultralong CDR3s and observed health traits.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"283 ","pages":"Article 110926"},"PeriodicalIF":1.4,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143724312","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}
Pub Date : 2025-04-01Epub Date: 2025-02-21DOI: 10.1016/j.vetimm.2025.110901
Jong Hyuk Kim , Donghee Lee , Kevin Hall , Hyunji Jo , John P. Bannantine , William C. Davis , Cleverson de Souza
Mycobacterium avium subspecies paratuberculosis (Map), the etiological agent of Johne’s disease in ruminants, poses challenges to veterinary health and food safety. Despite an immune response that partially controls early infection, Map persists in macrophages through mechanisms not well understood. Here, we explored how the Map major membrane protein (MMP) modulates immune pathways in bovine monocyte-derived macrophages (MoMΦs). MMP is a key component of the bacterial cell membrane recognized in cattle with Johne’s disease, making it a critical antigenic target for immune studies. Using high-resolution transcriptomics, we identified that MMP stimulation rapidly activates genes linked to pro-inflammatory cytokine signaling, antigen processing, and presentation via MHC I and II pathways. Gene Ontology and KEGG pathway enrichment analyses highlighted upregulation of TNF, IL-17, and NF-κB signaling cascades, suggesting an immune signaling that may foster cytotoxic T cell development. Phosphorylation assays confirmed that MMP triggers MAPK activation within minutes, implicating both p38 and JNK1/2 in early macrophage responses. Machine learning approaches revealed subtle yet significant MMP-specific gene signatures including ATG5 and ATG12, implicated in autophagosome assembly. These findings point to a dynamic interplay between antibacterial autophagy and immunostimulatory pathways elicited by MMP in bovine macrophages. Importantly, our results suggest the relevance of MMP as a potential vaccine target, as it not only elicits immune-activating signals but also engages host defenses critical to restricting Map survival. Overall, this work provides an ex vivo framework for delineating the molecular underpinnings of Map infection, offering new insights into macrophage-based immunity and informing development of novel therapeutic and prophylactic strategies against paratuberculosis. Our data open avenues for translational studies, illuminating the interplay between MMP, macrophages, and protective host immunity.
{"title":"Major membrane protein of Mycobacterium avium subp. paratuberculosis activates immune and autophagic pathways in bovine monocyte-derived macrophages","authors":"Jong Hyuk Kim , Donghee Lee , Kevin Hall , Hyunji Jo , John P. Bannantine , William C. Davis , Cleverson de Souza","doi":"10.1016/j.vetimm.2025.110901","DOIUrl":"10.1016/j.vetimm.2025.110901","url":null,"abstract":"<div><div><em>Mycobacterium avium</em> subspecies <em>paratuberculosis</em> (<em>Map</em>), the etiological agent of Johne’s disease in ruminants, poses challenges to veterinary health and food safety. Despite an immune response that partially controls early infection, <em>Map</em> persists in macrophages through mechanisms not well understood. Here, we explored how the <em>Map</em> major membrane protein (MMP) modulates immune pathways in bovine monocyte-derived macrophages (MoMΦs). MMP is a key component of the bacterial cell membrane recognized in cattle with Johne’s disease, making it a critical antigenic target for immune studies. Using high-resolution transcriptomics, we identified that MMP stimulation rapidly activates genes linked to pro-inflammatory cytokine signaling, antigen processing, and presentation via MHC I and II pathways. Gene Ontology and KEGG pathway enrichment analyses highlighted upregulation of TNF, IL-17, and NF-κB signaling cascades, suggesting an immune signaling that may foster cytotoxic T cell development. Phosphorylation assays confirmed that MMP triggers MAPK activation within minutes, implicating both p38 and JNK1/2 in early macrophage responses. Machine learning approaches revealed subtle yet significant MMP-specific gene signatures including <em>ATG5</em> and <em>ATG12</em>, implicated in autophagosome assembly. These findings point to a dynamic interplay between antibacterial autophagy and immunostimulatory pathways elicited by MMP in bovine macrophages. Importantly, our results suggest the relevance of MMP as a potential vaccine target, as it not only elicits immune-activating signals but also engages host defenses critical to restricting <em>Map</em> survival. Overall, this work provides an <em>ex vivo</em> framework for delineating the molecular underpinnings of <em>Map</em> infection, offering new insights into macrophage-based immunity and informing development of novel therapeutic and prophylactic strategies against paratuberculosis. Our data open avenues for translational studies, illuminating the interplay between MMP, macrophages, and protective host immunity.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"282 ","pages":"Article 110901"},"PeriodicalIF":1.4,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143510556","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}
扫码关注我们
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号