Veterinary immunology and immunopathology最新文献

{"title":"Pilot study: Understanding canine transmissible venereal tumor through its transcriptional profile","authors":"Paula de Sanctis Augusto ,&nbsp;Fernando Carmona Dinau ,&nbsp;Carlos Mario González-Zambrano ,&nbsp;Luis Mauricio Montoya-Flórez ,&nbsp;João Pessoa Araújo ,&nbsp;Noeme Sousa Rocha","doi":"10.1016/j.vetimm.2024.110818","DOIUrl":"10.1016/j.vetimm.2024.110818","url":null,"abstract":"<div><p>Canine transmissible venereal tumor (CTVT) is transmitted through the implantation of tumor cells. CTVT was the first tumor described with contagious characteristics and remains one of the few tumors with this capability. This study aimed to map the transcriptomic profile of CTVT to elucidate the potential mechanisms through which this tumor implants and evades host immune surveillance. For this study, 11 dogs aged ≥ 2 years diagnosed with CTVT were selected. Tumor biopsies were performed, RNA was extracted and converted into complementary DNA, followed by RT-qPCR analysis. The transcriptomic profile of CTVT revealed a wide array of differentially expressed genes. However, only the most relevant genes from an oncological perspective were discussed. IL-8, CXCL13, NCAM1, RNASEL, COROA1, and CBLB demonstrated potential associations with immune system evasion and transmission via implantation. Therefore, studying these genes may contribute to the development of targeted therapies that prevent contagion and immune evasion.</p></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"276 ","pages":"Article 110818"},"PeriodicalIF":1.4,"publicationDate":"2024-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142112460","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":"Codon optimization of voraxin α sequence enhances the immunogenicity of a recombinant vaccine against Hyalomma anatolicum infestation in rabbits","authors":"Zohre Monjezi ,&nbsp;Hedaiat allah Rooshanfekr ,&nbsp;Mahmood Nazari ,&nbsp;Fatemeh Salabi ,&nbsp;Mohammad Reza Tabandeh","doi":"10.1016/j.vetimm.2024.110817","DOIUrl":"10.1016/j.vetimm.2024.110817","url":null,"abstract":"<div><p>Research has shown that voraxin α derived from male ticks stimulates blood feeding to engorge in female ticks. Whereas, the oviposition rate, egg weight, and body weight of female ticks were reduced in animals vaccinated with recombinant (r-) voraxin α. These data suggest a potential role of r-voraxin α as a functional anti-tick antigen in <em>Rhipicephalus appendiculatus</em> and <em>Amblyomma hebraeum</em> tick infestation. This study investigated the immunogenicity of r-voraxin α protein from <em>Hyalomma anatolicum</em> (<em>H. anatolicum</em>) tick as an anti-tick vaccine in rabbits. The <em>H. anatolicum</em> voraxin α sequence was optimized according to the codon usage in E. coli before being sub-cloned into pQE30. The gene sequence of the voraxin α was synthesized, verified by DNA sequencing, cloned in a pQE30 vector, and transformed into E. coli. Then, the expression of the r-voraxin α protein was confirmed by SDS-PAGE and Western blot analysis. Subsequently, three rabbits were immunized with the r-voraxin α as the vaccinated group, whereas three rabbits without injection were considered the control group. The result indicated the success of cloning of codon-optimized <em>H. anatolicum</em> voraxin α gene. Moreover, the expression of the r-voraxin α protein (approximately 18 kDa) in the bacterial expression system was confirmed by SDS-PAGE and Western blot analysis. The results of this study showed that the mortality rate in vaccine recipients increased compared to the control group (<em>P &lt; 0.01</em>). Also, the egg weight, oviposition rate, and engorgement weight of female ticks fed from vaccinated animals were significantly reduced compared to the control group (<em>P &lt; 0.01</em>). The results confirmed that the codon-optimized <em>H. anatolicum</em> voraxin α gene expressed in the bacterial expression system could be a suitable anti-tick vaccine against <em>H. anatolicum</em> tick infestation.</p></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"275 ","pages":"Article 110817"},"PeriodicalIF":1.4,"publicationDate":"2024-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142084048","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 CD4+ T cell differentiation and diversity in dogs","authors":"Haeree P. Lang ,&nbsp;Kevin C. Osum ,&nbsp;Steven G. Friedenberg","doi":"10.1016/j.vetimm.2024.110816","DOIUrl":"10.1016/j.vetimm.2024.110816","url":null,"abstract":"<div><p>CD4⁺ T cells are an integral component of the adaptive immune response, carrying out many functions to combat a diverse range of pathogenic challenges. These cells exhibit remarkable plasticity, differentiating into specialized subsets such as T helper type 1 (T_H1), T_H2, T_H9, T_H17, T_H22, regulatory T cells (Tregs), and follicular T helper (T_FH) cells. Each subset is capable of addressing a distinct immunological need ranging from pathogen eradication to regulation of immune homeostasis. As the immune response subsides, CD4⁺ T cells rest down into long-lived memory phenotypes—including central memory (T_CM), effector memory (T_EM), resident memory (T_RM), and terminally differentiated effector memory cells (T_EMRA) that are localized to facilitate a swift and potent response upon antigen re-encounter. This capacity for long-term immunological memory and rapid reactivation upon secondary exposure highlights the role CD4⁺ T cells play in sustaining both adaptive defense mechanisms and maintenance.</p><p>Decades of mouse, human, and to a lesser extent, pig T cell research has provided the framework for understanding the role of CD4⁺ T cells in immune responses, but these model systems do not always mimic each other. Although our understanding of pig immunology is not as extensive as mouse or human research, we have gained valuable insight by studying this model. More akin to pigs, our understanding of CD4⁺ T cells in dogs is much less complete. This disparity exists in part because canine immunologists depend on paradigms from mouse and human studies to characterize CD4⁺ T cells in dogs, with a fraction of available lineage-defining antibody markers. Despite this, every major CD4⁺ T cell subset has been described to some extent in dogs. These subsets have been studied in various contexts, including <em>in vitro</em> stimulation, homeostatic conditions, and across a range of disease states. Canine CD4⁺ T cells have been categorized according to lineage-defining characteristics, trafficking patterns, and what cytokines they produce upon stimulation. This review addresses our current understanding of canine CD4⁺ T cells from a comparative perspective by highlighting both the similarities and differences from mouse, human, and pig CD4⁺ T cell biology. We also discuss knowledge gaps in our current understanding of CD4⁺ T cells in dogs that could provide direction for future studies in the field.</p></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"275 ","pages":"Article 110816"},"PeriodicalIF":1.4,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142037145","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":"Determination of systemic inflammation response index (SIRI), systemic inflammatory index (SII), HMGB1, Mx1 and TNF levels in neonatal calf diarrhea with systemic inflammatory response syndrome","authors":"Ömer Aydın ,&nbsp;Betül Apaydın Yıldırım","doi":"10.1016/j.vetimm.2024.110815","DOIUrl":"10.1016/j.vetimm.2024.110815","url":null,"abstract":"<div><p>The objective of this study was to examine the values of MX dynamin-like GTPase 1 (Mx1), high mobility group box-1 (HMGB1), systemic inflammatory response index (SIRI), systemic inflammatory index (SII), tumor necrosis factor (TNF), and other hematological indices in calves with systemic inflammatory response syndrome (SIRS). The study material was divided into two groups: the SIRS group (comprising 13 calves) and the control group (comprising 10 calves). The independent samples t-test and Mann-Whitney U test were employed for normally distributed and non-normally distributed data, respectively. The relationship between the two groups was determined using Spearman correlation coefficient analysis. Significant differences were identified between the SIRS group and the control group with regard to white blood cell (WBC; <em>P</em> &lt; 0.05), neutrophil (NEU; <em>P</em> &lt; 0.01), and neutrophil-to-lymphocyte ratio (NLR; <em>P</em> &lt; 0.001) values, in addition to SIRI (<em>P</em> &lt; 0.05), SII (<em>P</em> &lt; 0.01) values. Furthermore, HMGB1 (<em>P</em> &lt; 0.001), Mx1 (<em>P</em> &lt; 0.05), and TNF values (<em>P</em> &lt; 0.001) demonstrated notable disparities between the two groups. As a result of this study, it was concluded that there were significant increases in inflammatory hematological indices, as well as in the levels of HMGB1, Mx1, and TNF, in calves with SIRS.</p></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"275 ","pages":"Article 110815"},"PeriodicalIF":1.4,"publicationDate":"2024-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141992741","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":"Vaccination of cattle with a virus vector vaccine against a major membrane protein of Mycobacterium avium subsp. paratuberculosis elicits CD8 cytotoxic T cells that kill intracellular bacteria","authors":"Asmaa H. Mahmoud ,&nbsp;Gaber S. Abdellrazeq ,&nbsp;Valentina Franceschi ,&nbsp;David A. Schneider ,&nbsp;John P. Bannantine ,&nbsp;Lindsay M. Fry ,&nbsp;Victoria Hulubei ,&nbsp;Giovanna De Matteis ,&nbsp;Kun Taek Park ,&nbsp;Sergio Minesso ,&nbsp;William C. Davis ,&nbsp;Gaetano Donofrio","doi":"10.1016/j.vetimm.2024.110814","DOIUrl":"10.1016/j.vetimm.2024.110814","url":null,"abstract":"<div><p>Analysis of the recall response ex vivo in cattle vaccinated with a <em>Mycobacterium a</em>vium subsp. <em>paratuberculosis (Map) rel</em> deletion mutant revealed the immune response was directed toward a 35 kD major membrane protein (MMP) of <em>Map</em>. Antigen presenting cells (APC) primed with MMP elicited expansion of CD8 cytotoxic memory T cells (CTL) with ability to kill intracellular bacteria. Development of CTL was MHC-restricted. The gene <em>MAP2121c,</em> encoding MMP, was modified for expression of MMP (tPA-MMP-2mut) in a mammalian cell line to explore the potential of developing MMP as a vaccine. Ex vivo stimulation of PBMC, from <em>Map</em> free cattle, with APC primed with tPA-MMP-2mut expressed p35 elicited a primary CD8 CTL response comparable to the recall response elicited with PBMC from cattle vaccinated with either the <em>Maprel</em> deletion mutant or MMP. In the present study, the modified gene for MMP, now referred to as p35NN, was placed into a bovine herpes virus-4 (BoHV4) vector to determine the potential use of BoHV-4AΔTK-p35NN as a peptide-based vaccine. Subcutaneous vaccination of healthy cattle with BoHV-4AΔTK-p35NN elicited a CTL recall response, as detected ex vivo. The results show use of a virus vector is an effective way for delivery of MMP as a vaccine. The immunogenic activity of MMP was not lost when modified for expression in mammalian cells. The next step is to conduct a field trial to determine if presence of an immune response to MMP prevents <em>Map</em> from establishing an infection.</p></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"275 ","pages":"Article 110814"},"PeriodicalIF":1.4,"publicationDate":"2024-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141983352","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":"Expression of Toll-like receptors in Haemonchus Contortus resistant sheep: An innate immune parameter for host defense against gastrointestinal nematode infection","authors":"Rajiv Kumar,&nbsp;A.S. Meena,&nbsp;Trusha Baraiya,&nbsp;C.P. Swarnkar,&nbsp;S.S. Misra,&nbsp;Arun Kumar","doi":"10.1016/j.vetimm.2024.110813","DOIUrl":"10.1016/j.vetimm.2024.110813","url":null,"abstract":"<div><p>Innate immune parameters, a first line of defense against invading pathogens like bacteria, parasites, fungi, etc, play a significant role in the prevention and elimination of aetiological agents primarily by recognition of invading pathogen-specific molecules by different pattern recognition receptors. Toll-like receptors (TLRs), a type-I transmembrane glycoprotein, cause innate immune responses mainly by produing inflammatory cytokines, chemokines and interferons. The objective of present study was to determine the role of TLRs in parasite resistance in Malpura sheep. In the current study, transcript variation of TLRs and its downstream signalling molecules namely MyD88, TRIF, IRF-3, TRAF, TGF-β, NFκB, and CD14 were ascertained by real-time PCR in <em>Haemonchus contortus</em> resistant (R) and susceptible (S) Malpura sheep. Results have shown significantly (P&lt;0.05) up-regulated expression of TLR-2, TLR-4, TLR-5, TLR-8 and TLR-10 in July however down-regulated patterns were observed in August and September in R-line sheep compared to S-line sheep. This indicates that at more or less equal parasite load, the TLR genes in R sheep produce more transcripts, but after parasite loads have increased hugely in the S line, they easily surpass the levels seen in the S line. Result suggests that transcriptional activity of the TLR genes was related to parasite load and there were differences between the lines at different infection intensities. Three-point transcript expression observation of the signalling molecules namely TRIF, IRF-3, TRAF, a similar pattern was observed in R sheep compared with S sheep.</p></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"275 ","pages":"Article 110813"},"PeriodicalIF":1.4,"publicationDate":"2024-08-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141983351","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":"Newly developed peptide-ELISA successfully detected anti-IgG antibodies against Maedi-Visna virus in sheep","authors":"Ecem Su Koçkaya ,&nbsp;Hüseyin Can ,&nbsp;Yalçın Yaman ,&nbsp;Çağrı Kandemir ,&nbsp;Turgay Taşkın ,&nbsp;Muhammet Karakavuk ,&nbsp;Aysu Değirmenci Döşkaya ,&nbsp;Mert Döşkaya ,&nbsp;Erkan Pehlivan ,&nbsp;Halit Deniz Şireli ,&nbsp;Adnan Yüksel Gürüz ,&nbsp;Cemal Ün","doi":"10.1016/j.vetimm.2024.110806","DOIUrl":"10.1016/j.vetimm.2024.110806","url":null,"abstract":"<div><p>Maedi Visna Virus (MVV) is a retrovirus that can infect sheep. There is still no effective therapy or vaccine against this virus and timely diagnosis is important to combat the complications of the disease. In this study, we aimed to develop an ELISA using peptides derived from gag protein as antigen. For this purpose, B cell epitopes of gag protein were predicted and a docking analysis with the B cell receptor was performed to select peptides to be used in ELISA. After three soluble epitopes with the highest antigenicity were produced as peptides, the immunogenicity of each peptide was determined by ELISA using sheep serum samples categorized as MVV positive (n=24) and negative (n=13). Subsequently, <em>in house</em> ELISA using above mentioned immunogenic peptides as antigen was used to investigate MVV seroprevalence in sheep (n=88). According to the results, among three peptides, two of them strongly reacted with MVV positive serum samples and the mean absorbance values detected among positive and negative serum samples were statistically significant, indicating that these peptides were immunogenic (<em>P</em>=0.016 and <em>P</em>=0.038). The third peptide also reacted with positive serum samples but the mean absorbance value was not statistically significant and this peptide was considered non-immunogenic (<em>P</em>=0.175). The immunogenic two peptides showed the same high sensitivity and specificity values of 91.60 and 92.80 according to the commercial kit. Moreover, MVV seroprevalence detected by peptide-ELISAs using CKQGSKE and CRPQGKAGHKG peptides as antigen was 3.40 % and 4.5 %, respectively. As a result, it was shown that these peptides can be successfully used for serological diagnosis of MVV.</p></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"274 ","pages":"Article 110806"},"PeriodicalIF":1.4,"publicationDate":"2024-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141604236","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":"What we know about alterations in immune cells during sepsis in veterinary animals?","authors":"Izabela Siemińska ,&nbsp;Zbigniew Arent","doi":"10.1016/j.vetimm.2024.110804","DOIUrl":"10.1016/j.vetimm.2024.110804","url":null,"abstract":"<div><p>Sepsis is still one of the most common causes of death of animals and humans. It is marked by an aberrant immune response to infection, resulting in extensive inflammation, organ dysfunction, and, in severe instances, organ failure. Recognizable symptoms and markers of sepsis encompass substantial elevations in body temperature, respiratory rate, hemoglobin levels, and alterations in immune cell counts, including neutrophils, monocytes, and basophils, along with increases in certain acute-phase proteins. In contrast to human medicine, veterinarians must take into account some species differences. This article provides a comprehensive overview of changes in the immune system during sepsis, placing particular emphasis on species variations and exploring potential future drugs and interventions. Hence, understanding the intricate balance of the immune responses during sepsis is crucial to develop effective treatments and interventions to improve the chances of recovery in animals suffering from this serious condition.</p></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"274 ","pages":"Article 110804"},"PeriodicalIF":1.4,"publicationDate":"2024-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0165242724000904/pdfft?md5=1b28d7168d788e61c91461d3700fb20a&pid=1-s2.0-S0165242724000904-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141604237","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The mRNA vaccine platform for veterinary species","authors":"Fatemeh Fazel,&nbsp;Janan Shoja Doost,&nbsp;Sugandha Raj,&nbsp;Nitish Boodhoo,&nbsp;Khalil Karimi,&nbsp;Shayan Sharif","doi":"10.1016/j.vetimm.2024.110803","DOIUrl":"https://doi.org/10.1016/j.vetimm.2024.110803","url":null,"abstract":"<div><p>Vaccination has proven to be an effective means of controlling pathogens in animals. Since the introduction of veterinary vaccines in the 19th century, several generations of vaccines have been introduced. These vaccines have had a positive impact on global animal health and production. Despite, the success of veterinary vaccines, there are still some pathogens for which there are no effective vaccines available, such as African swine fever. Further, animal health is under the constant threat of emerging and re-emerging pathogens, some of which are zoonotic and can pose a threat to human health. The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has highlighted the need for new vaccine platforms that are safe and efficacious, but also importantly, are adaptable and can be modified rapidly to match the circulating pathogens. mRNA vaccines have been shown to be an effective vaccine platform against various viral and bacterial pathogens. This review will cover some of the recent advances in the field of mRNA vaccines for veterinary species. Moreover, various mRNA vaccines and their delivery methods, as well as their reported efficacy, will be discussed. Current limitations and future prospects of this vaccine platform in veterinary medicine will also be discussed.</p></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"274 ","pages":"Article 110803"},"PeriodicalIF":1.4,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0165242724000898/pdfft?md5=362a325dc3026508a40381bf985722be&pid=1-s2.0-S0165242724000898-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141605967","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Monoclonal antibodies for equine IL-1β enable the quantification of mature IL-1β in horses","authors":"Susanna Babasyan,&nbsp;Alicia Rollins,&nbsp;Bettina Wagner","doi":"10.1016/j.vetimm.2024.110805","DOIUrl":"10.1016/j.vetimm.2024.110805","url":null,"abstract":"<div><p>Interleukin-1β (IL-1β) is one of the key mediators of inflammation during innate immune responses. Mature bioactive IL-1β mediates essential host defense mechanisms but also has a mechanistic role in several autoinflammatory and degenerative diseases. In horses, specific and sensitive assays for IL-1β are crucial for immunological research on inflammatory processes and diseases. In this article, we describe the development of four monoclonal antibodies (mAbs) against equine IL-1β. The specificity of the new IL-1β mAbs was confirmed using a panel of equine recombinant cytokines and chemokines. The mAbs were validated for detection of native mature IL-1β in a fluorescent bead-based assay and for staining of IL-1β-producing immune cells by flow cytometry. The bead-based assay for equine IL-1β had a linear quantification range between 60<!--> <!-->pg/ml to 960<!--> <!-->ng/ml. Horse peripheral blood mononuclear cells (PBMC) secreted IL-1β after lipopolysaccharide (LPS) stimulation in time and dose dependent manner as quantified by the new equine IL-1β bead-based assay. A comparison of two commercial equine IL-1β ELISA kits with the new IL-1β fluorescent bead-based assay revealed that the bead-based assay improved the quantification of native equine IL-1β in LPS stimulated PBMC supernatants by detecting it with high intensity and a broad linear quantification range, while both ELISAs resulted in low signals and poor native IL-1β recognition. Intracellular staining and flow cytometric analysis confirmed that the main cellular source of IL-1β in equine PBMC after LPS stimulation were CD14⁺ monocytes. IL-1β secretion from PBMC was inhibited by a caspase inhibitor but protein translation within the cells was not, supporting the accumulation of pro-IL-1β within the cells even when proteolytic cleavage for IL-1β activation is missing. This confirmed the importance of specific mAbs for analyzing the biologically active, mature IL-1β in horses.</p></div>","PeriodicalId":23511,"journal":{"name":"Veterinary immunology and immunopathology","volume":"274 ","pages":"Article 110805"},"PeriodicalIF":1.4,"publicationDate":"2024-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0165242724000916/pdfft?md5=f91443a7738b5cf6e23890889a1aae98&pid=1-s2.0-S0165242724000916-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141604235","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}