Pub Date : 2025-06-13DOI: 10.1016/j.vetimm.2025.110965
Shreya Gopinath , Madhusudan Hosamani , Suresh Basagoudanavar , B.P. Sreenivasa , Biljo V. Joseph , Sharanagouda S. Patil
Classical swine fever (CSF), caused by the classical swine fever virus (CSFV; genus Pestivirus), is a highly transmissible disease in pigs that leads to severe economic losses for pig farmers worldwide. Early and accurate diagnosis of CSFV infections is crucial for effective containment of the disease, especially in endemic areas. In this study, the full-length viral protein Erns was successfully expressed in E. coli. The recombinant protein was purified using Ni-NTA affinity chromatography, and its antigenicity confirmed by western blot and indirect ELISA (iELISA) using CSFV-specific antiserum. The purified Erns protein was then used as an immunogen to generate murine hybridomas producing monoclonal antibodies (mAbs). A panel of 15 mAbs targeting this protein was characterized using iELISA and western blotting, for their ability to react with native antigens in cultures infected with CSFV. These mAbs have the potential to be used in the development of diagnostic tools, particularly for the detection and titration of virus-infected cultures through endpoint dilution assays.
{"title":"Monoclonal antibodies targeting the Erns protein of classical swine fever virus: application for virus detection","authors":"Shreya Gopinath , Madhusudan Hosamani , Suresh Basagoudanavar , B.P. Sreenivasa , Biljo V. Joseph , Sharanagouda S. Patil","doi":"10.1016/j.vetimm.2025.110965","DOIUrl":"10.1016/j.vetimm.2025.110965","url":null,"abstract":"<div><div>Classical swine fever (CSF), caused by the classical swine fever virus (CSFV; genus <em>Pestivirus</em>), is a highly transmissible disease in pigs that leads to severe economic losses for pig farmers worldwide. Early and accurate diagnosis of CSFV infections is crucial for effective containment of the disease, especially in endemic areas. In this study, the full-length viral protein E<sup>rns</sup> was successfully expressed in <em>E. coli</em>. The recombinant protein was purified using Ni-NTA affinity chromatography, and its antigenicity confirmed by western blot and indirect ELISA (iELISA) using CSFV-specific antiserum. The purified E<sup>rns</sup> protein was then used as an immunogen to generate murine hybridomas producing monoclonal antibodies (mAbs). A panel of 15 mAbs targeting this protein was characterized using iELISA and western blotting, for their ability to react with native antigens in cultures infected with CSFV. These mAbs have the potential to be used in the development of diagnostic tools, particularly for the detection and titration of virus-infected cultures through endpoint dilution assays.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"286 ","pages":"Article 110965"},"PeriodicalIF":1.4,"publicationDate":"2025-06-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144322926","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 : 2025-06-03DOI: 10.1016/j.vetimm.2025.110964
Muhammad Farooq , Awais Ghaffar , Ahmed Ali , Ryan Rahimi , Muhammad Azhar , Ishara M. Isham , Heshanthi Herath-Mudiyanselage , Sufna M. Suhail , Mohamed Faizal Abdul-Careem
This study investigated the impact of poor drinking water quality on infectious bronchitis virus (IBV) pathogenesis. Drinking water samples from Alberta layer farms were assessed based on physical, chemical, and microbiological properties. The highest-scoring field water (FW), which is suboptimal with higher pH, hardness and bicarbonate concentration was selected, transported in clean containers, and used in this control experiment. Forty-eight specific pathogen free White Leghorn chicks were divided into four groups: Tap water non-infected (TW-control), field water non-infected (FW-control), tap water infected (TW-infected), and field water infected (FW-infected). They were maintained on their respective water types for 7 weeks. The IBV genome load was significantly higher in the lungs of the FW-infected when compared to TW-infected group at 4 days post-infection (dpi). The histopathological lesion scores in the trachea and lungs were higher in the FW-infected birds when compared to the uninfected controls at observed time points. However, the histopathological lesion scores in the trachea and lungs of the TW-infected birds were not different when compared to that of FW-infected group. In the lungs, the CD4 + and CD8 + T cell populations were significantly higher in the TW-infected group at observed time points when compared to uninfected controls. However, the CD4 + and CD8 + T cell populations in lungs of the FW-infected birds were not different when compared to that of TW-infected group. In the spleen, CD4 + and CD8 + T cell populations were significantly higher in TW-infected and FW-infected birds when compared to uninfected controls depending on the observed time point and we did not observe differences in CD4 + and CD8 + T cell populations in spleen between TW-infected and FW-infected birds. These findings suggest that sub-optimal drinking water can exacerbate IBV infection by weakening immune responses and increasing disease severity. Further studies are necessary to observe the effect of suboptimal water quality on the development of vaccine-mediated immune response. Understanding these interactions is key for improving water management strategies for maintaining poultry health and productivity.
{"title":"Negative influence of suboptimal quality of drinking water on avian coronavirus pathogenesis and immune response: A Controlled Study","authors":"Muhammad Farooq , Awais Ghaffar , Ahmed Ali , Ryan Rahimi , Muhammad Azhar , Ishara M. Isham , Heshanthi Herath-Mudiyanselage , Sufna M. Suhail , Mohamed Faizal Abdul-Careem","doi":"10.1016/j.vetimm.2025.110964","DOIUrl":"10.1016/j.vetimm.2025.110964","url":null,"abstract":"<div><div>This study investigated the impact of poor drinking water quality on infectious bronchitis virus (IBV) pathogenesis. Drinking water samples from Alberta layer farms were assessed based on physical, chemical, and microbiological properties. The highest-scoring field water (FW), which is suboptimal with higher pH, hardness and bicarbonate concentration was selected, transported in clean containers, and used in this control experiment. Forty-eight specific pathogen free White Leghorn chicks were divided into four groups: Tap water non-infected (TW-control), field water non-infected (FW-control), tap water infected (TW-infected), and field water infected (FW-infected). They were maintained on their respective water types for 7 weeks. The IBV genome load was significantly higher in the lungs of the FW-infected when compared to TW-infected group at 4 days post-infection (dpi). The histopathological lesion scores in the trachea and lungs were higher in the FW-infected birds when compared to the uninfected controls at observed time points. However, the histopathological lesion scores in the trachea and lungs of the TW-infected birds were not different when compared to that of FW-infected group. In the lungs, the CD4 + and CD8 + T cell populations were significantly higher in the TW-infected group at observed time points when compared to uninfected controls. However, the CD4 + and CD8 + T cell populations in lungs of the FW-infected birds were not different when compared to that of TW-infected group. In the spleen, CD4 + and CD8 + T cell populations were significantly higher in TW-infected and FW-infected birds when compared to uninfected controls depending on the observed time point and we did not observe differences in CD4 + and CD8 + T cell populations in spleen between TW-infected and FW-infected birds. These findings suggest that sub-optimal drinking water can exacerbate IBV infection by weakening immune responses and increasing disease severity. Further studies are necessary to observe the effect of suboptimal water quality on the development of vaccine-mediated immune response. Understanding these interactions is key for improving water management strategies for maintaining poultry health and productivity.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"285 ","pages":"Article 110964"},"PeriodicalIF":1.4,"publicationDate":"2025-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144223624","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 : 2025-06-01DOI: 10.1016/j.vetimm.2025.110963
Emre Eren , Mustafa Sinan Aktaş
This study evaluated the immunomodulatory, antimicrobial, and anticoagulant effects of 1.25-dihydroxy vitamin D3 in neonatal calves with Escherichia coli-induced sepsis. Thirty neonatal Simmental calves were assigned to three groups: Control (n = 10), Medical Treatment (MT; n = 10), and Medical Treatment plus vitamin D3 (MT + D3; n = 10). The MT group received standard sepsis therapy, while the MT + D3 group was additionally administered 20,000 IU/kg intramuscular vitamin D3. Blood samples were collected on days 0, 1, 3, and 5 to analyze inflammatory cytokines (NF-κB, TNF-α, IL-1β, IL-10), cathelicidin, D-dimer, iron levels, and hematological parameters. Biochemical indicators including liver, kidney, and heart function, as well as calcium and vitamin D3 levels, were assessed on days 0 and 5. The MT + D3 group showed significant clinical and laboratory improvements compared to the MT group. Notably, SpO₂ levels increased, and metabolic acidosis resolved earlier. Hematological findings indicated reduced sepsis-associated anemia, with better preservation of RBC, HGB, and HCT levels. Inflammatory cytokines (NF-κB, TNF-α, IL-1β) significantly decreased, and IL-10 levels were more effectively regulated. Lower D-dimer levels indicated improved coagulation balance. Although cathelicidin levels initially increased, their subsequent decline by day 5 suggested controlled innate immune activation. In conclusion, vitamin D3 supplementation in combination with standard treatment effectively reduced systemic inflammation, supported innate immunity, and improved coagulation in neonatal calves with sepsis. These results suggest that vitamin D3 may serve as a beneficial adjunct therapy in bovine neonatal sepsis. Further research is needed to determine optimal dosing strategies and long-term clinical benefits.
{"title":"Effect of 1,25-dihydroxy vitamin D3 on inflammation, antimicrobial peptide, and D-dimer levels in Escherichia coli-induced sepsis in neonatal calves","authors":"Emre Eren , Mustafa Sinan Aktaş","doi":"10.1016/j.vetimm.2025.110963","DOIUrl":"10.1016/j.vetimm.2025.110963","url":null,"abstract":"<div><div>This study evaluated the immunomodulatory, antimicrobial, and anticoagulant effects of 1.25-dihydroxy vitamin D3 in neonatal calves with Escherichia coli-induced sepsis. Thirty neonatal Simmental calves were assigned to three groups: Control (<em>n</em> = 10), Medical Treatment (MT; <em>n</em> = 10), and Medical Treatment plus vitamin D<sub>3</sub> (MT + D<sub>3</sub>; <em>n</em> = 10). The MT group received standard sepsis therapy, while the MT + D<sub>3</sub> group was additionally administered 20,000 IU/kg intramuscular vitamin D<sub>3</sub>. Blood samples were collected on days 0, 1, 3, and 5 to analyze inflammatory cytokines (NF-κB, TNF-α, IL-1β, IL-10), cathelicidin, D-dimer, iron levels, and hematological parameters. Biochemical indicators including liver, kidney, and heart function, as well as calcium and vitamin D<sub>3</sub> levels, were assessed on days 0 and 5. The MT + D<sub>3</sub> group showed significant clinical and laboratory improvements compared to the MT group. Notably, SpO₂ levels increased, and metabolic acidosis resolved earlier. Hematological findings indicated reduced sepsis-associated anemia, with better preservation of RBC, HGB, and HCT levels. Inflammatory cytokines (NF-κB, TNF-α, IL-1β) significantly decreased, and IL-10 levels were more effectively regulated. Lower D-dimer levels indicated improved coagulation balance. Although cathelicidin levels initially increased, their subsequent decline by day 5 suggested controlled innate immune activation. In conclusion, vitamin D<sub>3</sub> supplementation in combination with standard treatment effectively reduced systemic inflammation, supported innate immunity, and improved coagulation in neonatal calves with sepsis. These results suggest that vitamin D<sub>3</sub> may serve as a beneficial adjunct therapy in bovine neonatal sepsis. Further research is needed to determine optimal dosing strategies and long-term clinical benefits.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"285 ","pages":"Article 110963"},"PeriodicalIF":1.4,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144195112","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 : 2025-05-31DOI: 10.1016/j.vetimm.2025.110962
Figen Celik , Muhammed Ahmed Selcuk , Muhammet Uslug , Sami Simsek
Echinococcus granulosus is a zoonotic helminth responsible for cystic echinococcosis, a significant public health concern. The diagnosis of E. granulosus infections in definitive hosts, such as dogs, is challenging due to the absence of clinical signs. This study aimed to evaluate the diagnostic performance of crude (EgSCA) and recombinant (rEgFN162) antigens for the detection of E. granulosus infection in dogs using ELISA and Western blot assays. Additionally, it sought to identify the most suitable antigen and method for population-based screening and post-treatment monitoring. Adult E. granulosus parasites were collected from experimentally infected dogs using arecoline hydrobromide purgation. Soluble crude antigen (EgSCA) was prepared through freeze-thaw cycles, sonication, and filtration, while recombinant fibronectin protein (rEgFN162) was obtained via gene cloning, expression, and purification in E. coli. The antigenic properties of EgSCA and rEgFN162 were analyzed by SDS-PAGE and Western blot. ELISA assays were performed to assess IgG and IgM responses in experimentally infected and treated dogs. Based on IgG ELISA results, EgSCA showed a sensitivity of 96.66 % and specificity of 66.66 %, while rEgFN162 demonstrated a sensitivity of 76.66 % and specificity of 46.66 %. In Western blot analysis, EgSCA achieved a sensitivity of 90 % and specificity of 83.33 %, whereas rEgFN162 showed 66.66 % sensitivity and 73.33 % specificity. The recombinant antigen showed a higher ability to differentiate E. granulosus infections from other helminth infections. The findings suggest that rEgFN162 is a promising candidate for the serodiagnosis of E. granulosus in dogs, with potential applications in epidemiological studies and post-treatment follow-up. Further validation with larger sample sizes is needed to confirm its diagnostic accuracy in natural infections.
{"title":"Serological diagnosis and post-treatment monitoring of Echinococcus granulosus in experimentally infected dogs using crude and recombinant fibronectin antigens","authors":"Figen Celik , Muhammed Ahmed Selcuk , Muhammet Uslug , Sami Simsek","doi":"10.1016/j.vetimm.2025.110962","DOIUrl":"10.1016/j.vetimm.2025.110962","url":null,"abstract":"<div><div><em>Echinococcus granulosus</em> is a zoonotic helminth responsible for cystic echinococcosis, a significant public health concern. The diagnosis of <em>E. granulosus</em> infections in definitive hosts, such as dogs, is challenging due to the absence of clinical signs. This study aimed to evaluate the diagnostic performance of crude (EgSCA) and recombinant (rEgFN162) antigens for the detection of <em>E. granulosus</em> infection in dogs using ELISA and Western blot assays. Additionally, it sought to identify the most suitable antigen and method for population-based screening and post-treatment monitoring. Adult <em>E. granulosus</em> parasites were collected from experimentally infected dogs using arecoline hydrobromide purgation. Soluble crude antigen (EgSCA) was prepared through freeze-thaw cycles, sonication, and filtration, while recombinant fibronectin protein (rEgFN162) was obtained via gene cloning, expression, and purification in <em>E. coli</em>. The antigenic properties of EgSCA and rEgFN162 were analyzed by SDS-PAGE and Western blot. ELISA assays were performed to assess IgG and IgM responses in experimentally infected and treated dogs. Based on IgG ELISA results, EgSCA showed a sensitivity of 96.66 % and specificity of 66.66 %, while rEgFN162 demonstrated a sensitivity of 76.66 % and specificity of 46.66 %. In Western blot analysis, EgSCA achieved a sensitivity of 90 % and specificity of 83.33 %, whereas rEgFN162 showed 66.66 % sensitivity and 73.33 % specificity. The recombinant antigen showed a higher ability to differentiate <em>E. granulosus</em> infections from other helminth infections. The findings suggest that rEgFN162 is a promising candidate for the serodiagnosis of <em>E. granulosus</em> in dogs, with potential applications in epidemiological studies and post-treatment follow-up. Further validation with larger sample sizes is needed to confirm its diagnostic accuracy in natural infections.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"285 ","pages":"Article 110962"},"PeriodicalIF":1.4,"publicationDate":"2025-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144195113","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 : 2025-05-31DOI: 10.1016/j.vetimm.2025.110961
Laís Sodré Santana Alves , Amanda Lopes Hasuda , Lucienne Garcia Pretto Giordano , Isabela Frederico , Isabelle Hadid dos Santos , Priscilla Fajardo Valente Pereira , Júlio Augusto Naylor Lisbôa , Karina Keller Marques da Costa Flaiban
This study investigated isolation of equine neutrophils and the assessment of their antioxidant function using the nitroblue tetrazolium (NBT) reduction test, an indicator of reactive oxygen species (ROS) activation. The objective was to test the viability of neutrophils isolated from equine whole blood and their ability to undergo respiratory burst, comparing the results with whole blood. The hypothesis tested was that isolation of equine neutrophils allows for effective functional evaluation, even after isolation process, with no significant differences between whole blood and isolated cells. The study showed that neutrophils maintained their ability to reduce NBT dye over a 6-hour period, with no significant differences between time points analyzed (0 h, 3 h, and 6 h). Isolation was performed using Histopaque®, and total neutrophil concentration and oxidative function were evaluated at three different time points. Cell counts and viability assessments were conducted using both automated and manual techniques, confirming the prolonged viability of neutrophils and the accuracy of antioxidant function testing.
{"title":"Does gradient isolation preserve equine neutrophil viability and oxidative function for up to six hours?","authors":"Laís Sodré Santana Alves , Amanda Lopes Hasuda , Lucienne Garcia Pretto Giordano , Isabela Frederico , Isabelle Hadid dos Santos , Priscilla Fajardo Valente Pereira , Júlio Augusto Naylor Lisbôa , Karina Keller Marques da Costa Flaiban","doi":"10.1016/j.vetimm.2025.110961","DOIUrl":"10.1016/j.vetimm.2025.110961","url":null,"abstract":"<div><div>This study investigated isolation of equine neutrophils and the assessment of their antioxidant function using the nitroblue tetrazolium (NBT) reduction test, an indicator of reactive oxygen species (ROS) activation. The objective was to test the viability of neutrophils isolated from equine whole blood and their ability to undergo respiratory burst, comparing the results with whole blood. The hypothesis tested was that isolation of equine neutrophils allows for effective functional evaluation, even after isolation process, with no significant differences between whole blood and isolated cells. The study showed that neutrophils maintained their ability to reduce NBT dye over a 6-hour period, with no significant differences between time points analyzed (0 h, 3 h, and 6 h). Isolation was performed using Histopaque®, and total neutrophil concentration and oxidative function were evaluated at three different time points. Cell counts and viability assessments were conducted using both automated and manual techniques, confirming the prolonged viability of neutrophils and the accuracy of antioxidant function testing.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"285 ","pages":"Article 110961"},"PeriodicalIF":1.4,"publicationDate":"2025-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144189565","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 : 2025-05-29DOI: 10.1016/j.vetimm.2025.110960
Emily C. Ashley , Walter Fuchs , Barbara G. Klupp , Dirk Werling , Simon P. Graham , Jane C. Edwards
Pseudorabies viruses (PrV), the causative agent of Aujeszky’s disease, continues to cause economic losses to pig producers across Southeast Asia. PrV is controlled by vaccination with live attenuated vaccines, such as the Bartha K61 strain, which has also shown promise as a viral vector. Despite the success of live attenuated PrV vaccines and their utility to be engineered as vaccine vectors, studies to understand the basis of their immunogenicity are scarce. Here, porcine bone marrow-derived dendritic cells (BMDC) were differentiated by culture with FLT3-L, generating eight myeloid cell populations differing in CADM1, CD172a, CD14, CD163 and CD11c expression, and included CADM1high conventional (c)DC and CD14+ DC. In vitro infection of BMDC with GFP-expressing PrV strains Bartha K61 and virulent Kaplan revealed a more rapid infection with Bartha K61. Compared to PrV Kaplan infection, there was also an increase in maturation marker expression (MHC class II and CD80/86) in both infected and bystander BMDC populations following Bartha K61 infection. This was accompanied by a concomitant increased cytokine response. IL-12 and TNF production associated with the cDC and CD14+ DC subsets, suggests that infection of these cells may be key to the potent immunogenicity associated with PrV Bartha K61 vaccination.
{"title":"Attenuated but not virulent pseudorabies virus activates porcine bone marrow-derived dendritic cells","authors":"Emily C. Ashley , Walter Fuchs , Barbara G. Klupp , Dirk Werling , Simon P. Graham , Jane C. Edwards","doi":"10.1016/j.vetimm.2025.110960","DOIUrl":"10.1016/j.vetimm.2025.110960","url":null,"abstract":"<div><div>Pseudorabies viruses (PrV), the causative agent of Aujeszky’s disease, continues to cause economic losses to pig producers across Southeast Asia. PrV is controlled by vaccination with live attenuated vaccines, such as the Bartha K61 strain, which has also shown promise as a viral vector. Despite the success of live attenuated PrV vaccines and their utility to be engineered as vaccine vectors, studies to understand the basis of their immunogenicity are scarce. Here, porcine bone marrow-derived dendritic cells (BMDC) were differentiated by culture with FLT3-L, generating eight myeloid cell populations differing in CADM1, CD172a, CD14, CD163 and CD11c expression, and included CADM1<sup>high</sup> conventional (c)DC and CD14<sup>+</sup> DC. In vitro infection of BMDC with GFP-expressing PrV strains Bartha K61 and virulent Kaplan revealed a more rapid infection with Bartha K61. Compared to PrV Kaplan infection, there was also an increase in maturation marker expression (MHC class II and CD80/86) in both infected and bystander BMDC populations following Bartha K61 infection. This was accompanied by a concomitant increased cytokine response. IL-12 and TNF production associated with the cDC and CD14<sup>+</sup> DC subsets, suggests that infection of these cells may be key to the potent immunogenicity associated with PrV Bartha K61 vaccination.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"285 ","pages":"Article 110960"},"PeriodicalIF":1.4,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144203406","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 : 2025-05-29DOI: 10.1016/j.vetimm.2025.110959
Murat Uzti̇mür, Aysu Ece Şengül, Cennet Nur Ünal
{"title":"Corrigendum to “Evaluation of serum serotonin as a biomarker of intestinal inflammation in calves” [Vet. Immunol. Immunopathol. 284 (2025) 110947]","authors":"Murat Uzti̇mür, Aysu Ece Şengül, Cennet Nur Ünal","doi":"10.1016/j.vetimm.2025.110959","DOIUrl":"10.1016/j.vetimm.2025.110959","url":null,"abstract":"","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"285 ","pages":"Article 110959"},"PeriodicalIF":1.4,"publicationDate":"2025-05-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144182411","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 : 2025-05-24DOI: 10.1016/j.vetimm.2025.110951
Xuan-ang Wang , Hong-xuan Li , Lan-Lan Zheng , Shi-jie Ma , Ping-Li Wang , Li Zhao , Hong-Ying Chen
Porcine epidemic diarrhea virus (PEDV) is a swine enteropathogenic coronavirus causing severe diarrhea and high mortality in neonatal piglets. Pigs of all ages are susceptible to PEDV, and the humoral immune response plays an important role in preventing PEDV infection. However, there is little information on monoclonal antibodies (mAbs) against PEDV derived from single B cells of pigs. In this study, we aimed to develop mAbs using antigen-specific single B cells from peripheral blood mononuclear cells (PBMCs) of pigs via fluorescence-activated cell sorting (FACS). Subsequently, the variable region genes of pig-derived mAbs were amplified and cloned into the plasmid pcDNA3.4 bearing the constant region gene of porcine-derived antibody. Pig-derived mAbs were expressed by transfecting the resultant antibody plasmids into HEK293F cells and validated using indirect Enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA), and Western blotting. The results showed 60 double-positive (antigen+ and IgG+) single B cells were obtained by flow sorting, of which 36 were positive for PEDV and 24 were positive for the N protein of PEDV. A total of 21 mAbs were expressed and purified. Indirect ELISA results showed that 20 bound specifically to PEDV, 19 recognized the N protein, and none reacted with S1D protein. Seven mAbs reacted with PEDV HN2021, as revealed by IFA. Western blotting showed that three N protein-specific mAbs identified linear epitopes, while the remaining 16 N protein-specific mAbs may recognize conformational epitopes. This study laid a foundation for the structural analysis of PEDV and the development of diagnostic reagents and antiviral drug.
{"title":"Development and identification of porcine monoclonal antibodies against PEDV from single B cells","authors":"Xuan-ang Wang , Hong-xuan Li , Lan-Lan Zheng , Shi-jie Ma , Ping-Li Wang , Li Zhao , Hong-Ying Chen","doi":"10.1016/j.vetimm.2025.110951","DOIUrl":"10.1016/j.vetimm.2025.110951","url":null,"abstract":"<div><div>Porcine epidemic diarrhea virus (PEDV) is a swine enteropathogenic coronavirus causing severe diarrhea and high mortality in neonatal piglets. Pigs of all ages are susceptible to PEDV, and the humoral immune response plays an important role in preventing PEDV infection. However, there is little information on monoclonal antibodies (mAbs) against PEDV derived from single B cells of pigs. In this study, we aimed to develop mAbs using antigen-specific single B cells from peripheral blood mononuclear cells (PBMCs) of pigs via fluorescence-activated cell sorting (FACS). Subsequently, the variable region genes of pig-derived mAbs were amplified and cloned into the plasmid pcDNA3.4 bearing the constant region gene of porcine-derived antibody. Pig-derived mAbs were expressed by transfecting the resultant antibody plasmids into HEK293F cells and validated using indirect Enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA), and Western blotting. The results showed 60 double-positive (antigen<sup>+</sup> and IgG<sup>+</sup>) single B cells were obtained by flow sorting, of which 36 were positive for PEDV and 24 were positive for the N protein of PEDV. A total of 21 mAbs were expressed and purified. Indirect ELISA results showed that 20 bound specifically to PEDV, 19 recognized the N protein, and none reacted with S1D protein. Seven mAbs reacted with PEDV HN2021, as revealed by IFA. Western blotting showed that three N protein-specific mAbs identified linear epitopes, while the remaining 16 N protein-specific mAbs may recognize conformational epitopes. This study laid a foundation for the structural analysis of PEDV and the development of diagnostic reagents and antiviral drug.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"285 ","pages":"Article 110951"},"PeriodicalIF":1.4,"publicationDate":"2025-05-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144134880","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 : 2025-05-20DOI: 10.1016/j.vetimm.2025.110949
Tianyu Han , Yan Jiang , Zhijun Liu , Lulu Wang , Yiding Liu , Shanshan Fei , Yu Yang , Tong Wang , Baiwen Guan , Mengran Cui , Qi Zhang , Haibin Wang , Guangliang Shi
Zearalenone (ZEA) is a mycotoxin that is immunotoxic and causes intestinal damage. Hyperoside (HYP) is a natural flavonol side with a wide range of sources and has a variety of pharmacological effects. The aim of this study is to investigate the effect and mechanism of HYP on ΖΕΑ-induced intestinal immunosuppression and intestinal injury in piglets. Histological and ultrastructural changes in the ileum of piglets were observed by H&E staining and transmission electron microscopy. The changes of intestinal microorganisms in the ileum of piglets were detected by 16S rRNA technology. Intestinal chemical barriers (MUC-1 and MUC-2), and physical barriers (β-Catenin, TJ-3, TJ-2, MYLK, Claudin2, and Claudin3) were measured by qRT-PCR. The intestinal immune barrier (sIg A) was detected by Elisa. Immune-related cytokines (TLR-4, IL-1β, IFN-γ, IL-18, IL-6, IL-17, IL-8, IL-25, and TNF-α) were detected by qRT-PCR. The content of ZEA in serum and ileum tissue was detected by Elisa. WB and qRT-PCR were used to detect ferroptosis related indicators (SLC7A11, Gpx4, FTH1, PTGS2, and ACSL4). Our results showed that HYP attenuated ZEA-induced tissue and ultrastructure damage and restored the richness and diversity of intestinal flora in the ileum of piglets. In addition, HYP also alleviated the accumulation of ZEA in the intestine and serum by restoring the chemical, physical and immunological barriers of the ileum. Moreover, HYP was found to attenuate ZEA-induced intestinal ferroptosis. Taken together, our study suggests that HYP can be used as an effective strategy to mitigate ZEA exposure-induced intestinal barrier damage and immune suppression in piglets.
{"title":"Hyperoside improves intestinal mucosal immunity against zearalenone-induced intestinal barrier damage by regulating intestinal flora","authors":"Tianyu Han , Yan Jiang , Zhijun Liu , Lulu Wang , Yiding Liu , Shanshan Fei , Yu Yang , Tong Wang , Baiwen Guan , Mengran Cui , Qi Zhang , Haibin Wang , Guangliang Shi","doi":"10.1016/j.vetimm.2025.110949","DOIUrl":"10.1016/j.vetimm.2025.110949","url":null,"abstract":"<div><div>Zearalenone (ZEA) is a mycotoxin that is immunotoxic and causes intestinal damage. Hyperoside (HYP) is a natural flavonol side with a wide range of sources and has a variety of pharmacological effects. The aim of this study is to investigate the effect and mechanism of HYP on ΖΕΑ-induced intestinal immunosuppression and intestinal injury in piglets. Histological and ultrastructural changes in the ileum of piglets were observed by H&E staining and transmission electron microscopy. The changes of intestinal microorganisms in the ileum of piglets were detected by 16S rRNA technology. Intestinal chemical barriers (MUC-1 and MUC-2), and physical barriers (β-Catenin, TJ-3, TJ-2, MYLK, Claudin2, and Claudin3) were measured by qRT-PCR. The intestinal immune barrier (sIg A) was detected by Elisa. Immune-related cytokines (TLR-4, IL-1β, IFN-γ, IL-18, IL-6, IL-17, IL-8, IL-25, and TNF-α) were detected by qRT-PCR. The content of ZEA in serum and ileum tissue was detected by Elisa. WB and qRT-PCR were used to detect ferroptosis related indicators (SLC7A11, Gpx4, FTH1, PTGS2, and ACSL4). Our results showed that HYP attenuated ZEA-induced tissue and ultrastructure damage and restored the richness and diversity of intestinal flora in the ileum of piglets. In addition, HYP also alleviated the accumulation of ZEA in the intestine and serum by restoring the chemical, physical and immunological barriers of the ileum. Moreover, HYP was found to attenuate ZEA-induced intestinal ferroptosis. Taken together, our study suggests that HYP can be used as an effective strategy to mitigate ZEA exposure-induced intestinal barrier damage and immune suppression in piglets.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"285 ","pages":"Article 110949"},"PeriodicalIF":1.4,"publicationDate":"2025-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144166496","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 : 2025-05-19DOI: 10.1016/j.vetimm.2025.110950
Maysa Serpa Gonçalves, Elaine Maria Seles Dorneles
Brucella abortus exhibits the dissociation phenomenon, in which naturally smooth samples lose the O chain of lipopolysaccharide (LPS) and become rough, associated with changes in colony shape, culture characteristics, cell morphology, immunological reactions, biochemical reactions and, possibly, virulence. However, the significance and impact of S-R dissociation in cultures (in vitro) or even in vivo is unclear, especially considering that rough samples have already been isolated from clinical samples in different hosts and, also, are successfully used as vaccine strains. Thus, the objective of this study was to review the literature on Brucella spp. LPS to better understand the impact of the LPS morphology in B. abortus in the vaccinal efficacy. The available information indicates that is undeniable that LPS is related to virulence modulation and inducing immunity in the natural hosts of Brucella spp. However, the continuous emergence of rough variants in vivo (infection) or in vitro (cultivation of the microorganism) suggests that this phenotype is part of the biology of the agent and may confer some survival advantage to the bacteria. In fact, for some samples, the permanent or temporary loss of the O chain (O-PS), whether natural or induced, did not necessarily imply a decrease in virulence, immunogenicity, or post-challenge induced protection, since results in both directions were observed in the literature, depending mainly on the parental samples used and the silenced genes. Thus, it is concluded that the emergence of variants related to the smooth/rough LPS of a sample of B. abortus does not necessarily imply changes in the virulence/immunogenicity of that sample and, consequently, in vaccine potency or efficacy, in case of vaccine strains.
{"title":"Does lipopolysaccharide morphology (smooth or rough) of Brucella abortus vaccine strains influence the potency or efficacy of the vaccine?","authors":"Maysa Serpa Gonçalves, Elaine Maria Seles Dorneles","doi":"10.1016/j.vetimm.2025.110950","DOIUrl":"10.1016/j.vetimm.2025.110950","url":null,"abstract":"<div><div><em>Brucella abortus</em> exhibits the dissociation phenomenon, in which naturally smooth samples lose the O chain of lipopolysaccharide (LPS) and become rough, associated with changes in colony shape, culture characteristics, cell morphology, immunological reactions, biochemical reactions and, possibly, virulence. However, the significance and impact of S-R dissociation in cultures (<em>in vitro</em>) or even <em>in vivo</em> is unclear, especially considering that rough samples have already been isolated from clinical samples in different hosts and, also, are successfully used as vaccine strains. Thus, the objective of this study was to review the literature on <em>Brucella</em> spp. LPS to better understand the impact of the LPS morphology in <em>B. abortus</em> in the vaccinal efficacy. The available information indicates that is undeniable that LPS is related to virulence modulation and inducing immunity in the natural hosts of <em>Brucella</em> spp. However, the continuous emergence of rough variants <em>in vivo</em> (infection) or <em>in vitro</em> (cultivation of the microorganism) suggests that this phenotype is part of the biology of the agent and may confer some survival advantage to the bacteria. In fact, for some samples, the permanent or temporary loss of the O chain (O-PS), whether natural or induced, did not necessarily imply a decrease in virulence, immunogenicity, or post-challenge induced protection, since results in both directions were observed in the literature, depending mainly on the parental samples used and the silenced genes. Thus, it is concluded that the emergence of variants related to the smooth/rough LPS of a sample of <em>B. abortus</em> does not necessarily imply changes in the virulence/immunogenicity of that sample and, consequently, in vaccine potency or efficacy, in case of vaccine strains.</div></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"285 ","pages":"Article 110950"},"PeriodicalIF":1.4,"publicationDate":"2025-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144124370","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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