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1.
Front Immunol ; 15: 1368545, 2024.
Article in English | MEDLINE | ID: mdl-38835764

ABSTRACT

There is a rapidly growing interest in how the avian intestine is affected by dietary components and feed additives. The paucity of physiologically relevant models has limited research in this field of poultry gut health and led to an over-reliance on the use of live birds for experiments. The development of complex 3D intestinal organoids or "mini-guts" has created ample opportunities for poultry research in this field. A major advantage of the floating chicken intestinal organoids is the combination of a complex cell system with an easily accessible apical-out orientation grown in a simple culture medium without an extracellular matrix. The objective was to investigate the impact of a commercial proprietary blend of organic acids and essential oils (OA+EO) on the innate immune responses and kinome of chicken intestinal organoids in a Salmonella challenge model. To mimic the in vivo prolonged exposure of the intestine to the product, the intestinal organoids were treated for 2 days with 0.5 or 0.25 mg/mL OA+EO and either uninfected or infected with Salmonella and bacterial load in the organoids was quantified at 3 hours post infection. The bacteria were also treated with OA+EO for 1 day prior to challenge of the organoids to mimic intestinal exposure. The treatment of the organoids with OA+EO resulted in a significant decrease in the bacterial load compared to untreated infected organoids. The expression of 88 innate immune genes was investigated using a high throughput qPCR array, measuring the expression of 88 innate immune genes. Salmonella invasion of the untreated intestinal organoids resulted in a significant increase in the expression of inflammatory cytokine and chemokines as well as genes involved in intracellular signaling. In contrast, when the organoids were treated with OA+EO and challenged with Salmonella, the inflammatory responses were significantly downregulated. The kinome array data suggested decreased phosphorylation elicited by the OA+EO with Salmonella in agreement with the gene expression data sets. This study demonstrates that the in vitro chicken intestinal organoids are a new tool to measure the effect of the feed additives in a bacterial challenge model by measuring innate immune and protein kinases responses.


Subject(s)
Animal Feed , Chickens , Intestines , Organoids , Animals , Intestines/immunology , Intestines/drug effects , Intestines/microbiology , Immunity, Innate , Oils, Volatile/pharmacology , Salmonella Infections, Animal/immunology , Salmonella Infections, Animal/microbiology , Poultry Diseases/microbiology , Poultry Diseases/immunology , Intestinal Mucosa/microbiology , Intestinal Mucosa/immunology , Intestinal Mucosa/drug effects
2.
Vet Immunol Immunopathol ; 273: 110791, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38824909

ABSTRACT

Infectious bronchitis virus (IBV) strains of the Delmarva (DMV)/1639 genotype have been causing false layer syndrome (FLS) in the Eastern Canadian layer operations since the end of 2015. FLS is characterized by the development of cystic oviducts in layer pullets infected at an early age. Currently, there are no homologous vaccines for the control of this IBV genotype. Our previous research showed that a heterologous vaccination regimen incorporating Massachusetts (Mass) and Connecticut (Conn) IBV types protects layers against DMV/1639 genotype IBV. The aim of this study was to investigate the role of maternal antibodies conferred by breeders received the same vaccination regimen in the protection against the development of DMV/1639-induced FLS in pullets. Maternal antibody-positive (MA+) and maternal antibody-negative (MA-) female progeny chicks were challenged at 1 day of age and kept under observation for 16 weeks. Oviductal cystic formations were observed in 3 of 14 birds (21.4 %) in the MA- pullets, while the lesions were notably absent in the MA+ pullets. Milder histopathological lesions were observed in the examined tissues of the MA+ pullets. However, the maternal derived immunity failed to demonstrate protection against the damage to the tracheal ciliary activity, viral shedding, and viral tissue distribution. Overall, this study underscores the limitations of maternal derived immunity in preventing certain aspects of viral pathogenesis, emphasizing the need for comprehensive strategies to address different aspects of IBV infection.


Subject(s)
Antibodies, Viral , Chickens , Coronavirus Infections , Infectious bronchitis virus , Poultry Diseases , Viral Vaccines , Animals , Infectious bronchitis virus/immunology , Poultry Diseases/prevention & control , Poultry Diseases/immunology , Poultry Diseases/virology , Chickens/immunology , Chickens/virology , Female , Antibodies, Viral/blood , Antibodies, Viral/immunology , Viral Vaccines/immunology , Viral Vaccines/administration & dosage , Coronavirus Infections/prevention & control , Coronavirus Infections/veterinary , Coronavirus Infections/immunology , Coronavirus Infections/virology , Immunity, Maternally-Acquired , Trachea/immunology , Trachea/virology , Oviducts/immunology , Oviducts/pathology , Oviducts/virology
3.
PLoS Pathog ; 20(5): e1012261, 2024 May.
Article in English | MEDLINE | ID: mdl-38805555

ABSTRACT

Marek's disease virus (MDV) vaccines were the first vaccines that protected against cancer. The avirulent turkey herpesvirus (HVT) was widely employed and protected billions of chickens from a deadly MDV infection. It is also among the most common vaccine vectors providing protection against a plethora of pathogens. HVT establishes latency in T-cells, allowing the vaccine virus to persist in the host for life. Intriguingly, the HVT genome contains telomeric repeat arrays (TMRs) at both ends; however, their role in the HVT life cycle remains elusive. We have previously shown that similar TMRs in the MDV genome facilitate its integration into host telomeres, which ensures efficient maintenance of the virus genome during latency and tumorigenesis. In this study, we investigated the role of the TMRs in HVT genome integration, latency, and reactivation in vitro and in vivo. Additionally, we examined HVT infection of feather follicles. We generated an HVT mutant lacking both TMRs (vΔTMR) that efficiently replicated in cell culture. We could demonstrate that wild type HVT integrates at the ends of chromosomes containing the telomeres in T-cells, while integration was severely impaired in the absence of the TMRs. To assess the role of TMRs in vivo, we infected one-day-old chickens with HVT or vΔTMR. vΔTMR loads were significantly reduced in the blood and hardly any virus was transported to the feather follicle epithelium where the virus is commonly shed. Strikingly, latency in the spleen and reactivation of the virus were severely impaired in the absence of the TMRs, indicating that the TMRs are crucial for the establishment of latency and reactivation of HVT. Our findings revealed that the TMRs facilitate integration of the HVT genome into host chromosomes, which ensures efficient persistence in the host, reactivation, and transport of the virus to the skin.


Subject(s)
Chickens , Marek Disease , Telomere , Virus Integration , Virus Latency , Animals , Chickens/virology , Telomere/genetics , Telomere/virology , Marek Disease/virology , Marek Disease/immunology , Marek Disease/prevention & control , Genetic Vectors , Herpesvirus 1, Meleagrid/genetics , Herpesvirus 1, Meleagrid/immunology , Marek Disease Vaccines/immunology , Marek Disease Vaccines/genetics , Genome, Viral , Herpesvirus 2, Gallid/genetics , Herpesvirus 2, Gallid/immunology , Repetitive Sequences, Nucleic Acid , Poultry Diseases/virology , Poultry Diseases/immunology , Poultry Diseases/prevention & control
4.
Poult Sci ; 103(6): 103716, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38703453

ABSTRACT

Coccidiosis, a protozoan disease that substantially impacts poultry production, is characterized by an intracellular parasite. The study utilized 48 one-day-old Horro chickens, randomly divided into the infected (I) and control (C) groups. The challenge group of chickens were administered Eimeria maxima oocysts via oral gavage at 21-days-old, and each chicken received 2 mL containing 7×104 sporulated oocysts. The total RNAs of chicken jejunum and cecum tissues were isolated from three samples, each from I and C groups. Our study aimed to understand the host immune-parasite interactions and compare immune response mRNA profiles in chicken jejunum and cecum tissues at 4 and 7 days postinfection with Eimeria maxima. The results showed that 823 up- and 737 down-regulated differentially expressed mRNAs (DEmRNAs) in jejunum at 4 d infection and control (J4I vs. J4C), and 710 up- and 368 down-regulated DEmRNAs in jejunum at 7 days infection and control (J7I vs. J7C) were identified. In addition, DEmRNAs in cecum tissue, 1424 up- and 1930 down-regulated genes in cecum at 4 days infection and control (C4I vs. C4C), and 77 up- and 191 down-regulated genes in cecum at 7 days infection and control (C7I vs. C7C) were detected. The crucial DEmRNAs, including SLC7A5, IL1R2, GLDC, ITGB6, ADAMTS4, IL1RAP, TNFRSF11B, IMPG2, WNT9A, and FOXF1, played pivotal roles in the immune response during Eimeria maxima infection of chicken jejunum. In addition, the potential detection of FSTL3, RBP7, CCL20, DPP4, PRKG2, TFPI2, and CDKN1A in the cecum during the host immune response against Eimeria maxima infection is particularly noteworthy. Furthermore, our functional enrichment analysis revealed the primary involvement of DEmRNAs in small molecule metabolic process, immune response function, inflammatory response, and toll-like receptor 10 signaling pathway in the jejunum at 4 and 7 days postinfection. Similarly, in the cecum, DEmRNAs at 4 and 7 days postinfection were enriched in processes related to oxidative stress response and immune responses. Our findings provide new insights and contribute significantly to the field of poultry production and parasitology.


Subject(s)
Cecum , Chickens , Coccidiosis , Eimeria , Jejunum , Poultry Diseases , RNA, Messenger , Animals , Eimeria/physiology , Coccidiosis/veterinary , Coccidiosis/parasitology , Coccidiosis/immunology , Cecum/parasitology , Cecum/metabolism , Poultry Diseases/parasitology , Poultry Diseases/genetics , Poultry Diseases/metabolism , Poultry Diseases/immunology , Jejunum/parasitology , Jejunum/metabolism , RNA, Messenger/metabolism , RNA, Messenger/genetics , Transcriptome , Random Allocation
5.
Res Vet Sci ; 174: 105293, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38754221

ABSTRACT

Recently, several attempts have been made to replace egg-based with cell-based vaccines to prevent and control Infectious Bursal Disease Virus (IBDV). This study aimed to evaluate a new fish cell line (M99) for culturing and replicating IBDV. After observing complete cytopathic effects (CPE) on the M99 cell line, virus titers were determined using the TCID50 test, and the presence of the virus was confirmed using an RT-PCR test. Subsequently, 135 broiler chickens (14 days old) were randomly divided into three equal groups for immune response measurements: G1: immunized with a commercial vaccine, G2: immunized with an experimental vaccine, and G3: control. Antibody responses, bursal index, and histopathological evaluations were examined on different days after immunization. Based on the results, CPE of the virus was noticeable from the first passage, becoming complete by the third passage. The infectious titer of the virus was log106.9. Antibody titer measured 21 days after immunization in both vaccinated groups were significantly differed from the control group (p < 0.05). The results obtained from examining the bursal index and histopathological evaluations showed no significant difference between the studied groups at different times. Overall, this research is the first report on the successful cultivation of infectious bursal virus on a permanent cell line of fish origin, with the advantages of tolerance to a wide temperature range (26-40 degrees Celsius). Therefore, this cell line has potential for use to attenuate, cultivate, and adapt other pathogens to cold temperatures in future studies.


Subject(s)
Birnaviridae Infections , Chickens , Infectious bursal disease virus , Poultry Diseases , Viral Vaccines , Virus Replication , Infectious bursal disease virus/immunology , Animals , Viral Vaccines/immunology , Chickens/virology , Birnaviridae Infections/veterinary , Birnaviridae Infections/virology , Birnaviridae Infections/prevention & control , Birnaviridae Infections/immunology , Cell Line , Poultry Diseases/virology , Poultry Diseases/prevention & control , Poultry Diseases/immunology , Fishes/virology
6.
Vet Microbiol ; 294: 110119, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38772075

ABSTRACT

Mycoplasma synoviae causes infectious synovitis and respiratory tract infections in chickens and is responsible for significant economic losses in the poultry industry. Effective attachment and colonisation of the trachea is critical for the persistence of the organism and progression of the disease it causes. The respiratory tract infection is usually sub-clinical, but concurrent infection with infectious bronchitis virus (IBV) is known to enhance the pathogenicity of M. synoviae. This study aimed to explore differentially expressed genes in the tracheal mucosa, and their functional categories, during chronic infection with M. synoviae, using a M. synoviae-IBV infection model. The transcriptional profiles of the trachea were assessed 2 weeks after infection using RNA sequencing. In chickens infected with M. synoviae or IBV, only 1 or 8 genes were differentially expressed compared to uninfected chickens, respectively. In contrast, the M. synoviae-IBV infected chickens had 621 upregulated and 206 downregulated genes compared to uninfected chickens. Upregulated genes and their functional categories were suggestive of uncontrolled lymphoid cell proliferation and an ongoing pro-inflammatory response. Genes associated with anti-inflammatory effects, pathogen removal, apoptosis, regulation of the immune response, airway homoeostasis, cell adhesion and tissue regeneration were downregulated. Overall, transcriptional changes in the trachea, 2 weeks after infection with M. synoviae and IBV, indicate immune dysregulation, robust inflammation and a lack of cytotoxic damage during chronic infection. This model provides insights into the pathogenesis of chronic infection with M. synoviae.


Subject(s)
Chickens , Mycoplasma Infections , Mycoplasma synoviae , Poultry Diseases , Trachea , Animals , Mycoplasma Infections/veterinary , Mycoplasma Infections/microbiology , Mycoplasma Infections/immunology , Poultry Diseases/microbiology , Poultry Diseases/virology , Poultry Diseases/immunology , Mycoplasma synoviae/genetics , Trachea/microbiology , Trachea/virology , Infectious bronchitis virus/genetics , Infectious bronchitis virus/immunology , Infectious bronchitis virus/physiology , Chronic Disease , Coronavirus Infections/veterinary , Coronavirus Infections/virology , Coronavirus Infections/immunology , Transcriptome , Gene Expression Profiling , Coinfection/veterinary , Coinfection/microbiology , Coinfection/virology
7.
Microb Pathog ; 192: 106691, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38759933

ABSTRACT

Necrotic enteritis (NE) is a potentially fatal poultry disease that causes enormous economic losses in the poultry industry worldwide. The study aimed to evaluate the effects of dietary organic yeast-derived selenium (Se) on immune protection against experimental necrotic enteritis (NE) in commercial broilers. Chickens were fed basal diets supplemented with different Se levels (0.25, 0.50, and 1.00 Se mg/kg). To induce NE, Clostridium perfringens (C. perfringens) was orally administered at 14 days of age post hatch. The results showed that birds fed 0.25 Se mg/kg exhibited significantly increased body weight gain compared with the non-supplemented/infected birds. There were no significant differences in gut lesions between the Se-supplemented groups and the non-supplemented group. The antibody levels against α-toxin and NetB toxin increased with the increase between 0.25 Se mg/kg and 0.50 Se mg/kg. In the jejunal scrapings and spleen, the Se-supplementation groups up-regulated the transcripts for pro-inflammatory cytokines IL-1ß, IL-6, IL-8, iNOS, and LITAF and avian ß-defensin 6, 8, and 13 (AvBD6, 8 and 13). In conclusion, supplementation with organic yeast-derived Se alleviates the negative consequences and provides beneficial protection against experimental NE.


Subject(s)
Animal Feed , Chickens , Clostridium Infections , Clostridium perfringens , Cytokines , Dietary Supplements , Enteritis , Poultry Diseases , Selenium , Animals , Enteritis/prevention & control , Enteritis/veterinary , Enteritis/immunology , Enteritis/microbiology , Selenium/pharmacology , Selenium/administration & dosage , Poultry Diseases/prevention & control , Poultry Diseases/immunology , Clostridium perfringens/immunology , Clostridium Infections/prevention & control , Clostridium Infections/veterinary , Clostridium Infections/immunology , Cytokines/metabolism , Bacterial Toxins/immunology , Necrosis , beta-Defensins/metabolism , Jejunum/drug effects , Jejunum/immunology , Jejunum/microbiology , Jejunum/pathology , Spleen/immunology , Yeasts , Nitric Oxide Synthase Type II/metabolism , Interleukin-6/metabolism , Interleukin-8/metabolism , Interleukin-1beta/metabolism , Antibodies, Bacterial/blood
8.
Vet Microbiol ; 294: 110108, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38729093

ABSTRACT

H7N9 subtype avian influenza virus (AIV) poses a great challenge to poultry industry. Virus-like particle (VLP) is a prospective alternative for the traditional egg-based influenza vaccines. N-linked glycosylation (NLG) regulates the efficacy of influenza vaccines, whereas the impact of NLG modifications on the efficacy of influenza VLP vaccines remains unclear. Here, H7N9 VLPs were assembled in insect cells through co-infection with the baculoviruses expressing the NLG-modified hemagglutinin (HA), neuraminidase and matrix proteins, and the VLP vaccines were assessed in chickens and mice. NLG modifications significantly enhanced hemagglutination-inhibition and virus neutralization antibody responses in mice, rather than in chickens, because different immunization strategies were used in these animal models. The presence of dual NLG at residues 133 and 158 significantly elevated HA-binding IgG titers in chickens and mice. The VLP vaccines conferred complete protection and significantly suppressed virus replication and lung pathology post challenge with H7N9 viruses in chickens and mice. VLP immunization activated T cell immunity-related cytokine response and inhibited inflammatory cytokine response in mouse lung. Of note, the presence of dual NLG at residues 133 and 158 optimized the capacity of the VLP vaccine to stimulate interleukin-4 expression, inhibit virus shedding or alleviate lung pathology in chickens or mice. Intriguingly, the VLP vaccine with NLG addition at residue 133 provided partial cross-protection against the H5Nx subtype AIVs in chickens and mice. In conclusion, dual NLG at residues 133 and 158 in HA can be potentially used to enhance the efficacy of H7N9 VLP vaccines in chickens and mammals.


Subject(s)
Antibodies, Viral , Chickens , Hemagglutinin Glycoproteins, Influenza Virus , Influenza A Virus, H7N9 Subtype , Influenza Vaccines , Influenza in Birds , Mice, Inbred BALB C , Vaccines, Virus-Like Particle , Animals , Chickens/immunology , Influenza Vaccines/immunology , Influenza Vaccines/administration & dosage , Mice , Influenza A Virus, H7N9 Subtype/immunology , Vaccines, Virus-Like Particle/immunology , Vaccines, Virus-Like Particle/administration & dosage , Glycosylation , Influenza in Birds/prevention & control , Influenza in Birds/immunology , Influenza in Birds/virology , Antibodies, Viral/blood , Hemagglutinin Glycoproteins, Influenza Virus/immunology , Female , Orthomyxoviridae Infections/prevention & control , Orthomyxoviridae Infections/veterinary , Orthomyxoviridae Infections/immunology , Antibodies, Neutralizing/blood , Antibodies, Neutralizing/immunology , Cytokines , Poultry Diseases/prevention & control , Poultry Diseases/virology , Poultry Diseases/immunology
9.
Vaccine ; 42(18): 3756-3767, 2024 Jul 11.
Article in English | MEDLINE | ID: mdl-38724417

ABSTRACT

A Newcastle disease virus (NDV)-vectored vaccine expressing clade 2.3.4.4b H5 Hemagglutinin was developed and assessed for efficacy against H5N1 highly pathogenic avian influenza (HPAI) in specific pathogen-free (SPF) chickens, broilers, and domestic ducks. In SPF chickens, the live recombinant NDV-vectored vaccine, rK148/22-H5, achieved complete survival against HPAI and NDV challenges and significantly reduced viral shedding. Notably, the live rK148/22-H5 vaccine conferred good clinical protection in broilers despite the presence of maternally derived antibodies. Good clinical protection was observed in domestic ducks, with decreased viral shedding. It demonstrated complete survival and reduced cloacal viral shedding when used as an inactivated vaccine from SPF chickens. The rK148/22-H5 vaccine is potentially a viable and supportive option for biosecurity measure, effectively protecting in chickens against the deadly clade 2.3.4.4b H5 HPAI and NDV infections. Furthermore, it aligns with the strategy of Differentiating Infected from Vaccinated Animals (DIVA).


Subject(s)
Antibodies, Viral , Chickens , Ducks , Hemagglutinin Glycoproteins, Influenza Virus , Influenza A Virus, H5N1 Subtype , Influenza in Birds , Newcastle disease virus , Vaccines, Inactivated , Vaccines, Synthetic , Virus Shedding , Animals , Chickens/immunology , Influenza in Birds/prevention & control , Influenza in Birds/immunology , Newcastle disease virus/immunology , Newcastle disease virus/genetics , Influenza A Virus, H5N1 Subtype/immunology , Influenza A Virus, H5N1 Subtype/genetics , Influenza A Virus, H5N1 Subtype/pathogenicity , Ducks/virology , Ducks/immunology , Vaccines, Inactivated/immunology , Vaccines, Inactivated/administration & dosage , Vaccines, Synthetic/immunology , Vaccines, Synthetic/administration & dosage , Vaccines, Synthetic/genetics , Antibodies, Viral/immunology , Antibodies, Viral/blood , Hemagglutinin Glycoproteins, Influenza Virus/immunology , Hemagglutinin Glycoproteins, Influenza Virus/genetics , Influenza Vaccines/immunology , Influenza Vaccines/administration & dosage , Influenza Vaccines/genetics , Specific Pathogen-Free Organisms , Vaccines, Attenuated/immunology , Vaccines, Attenuated/administration & dosage , Vaccines, Attenuated/genetics , Poultry Diseases/prevention & control , Poultry Diseases/virology , Poultry Diseases/immunology , Newcastle Disease/prevention & control , Newcastle Disease/immunology , Viral Vaccines/immunology , Viral Vaccines/administration & dosage , Viral Vaccines/genetics
10.
PLoS Pathog ; 20(5): e1012232, 2024 May.
Article in English | MEDLINE | ID: mdl-38743760

ABSTRACT

Infectious bronchitis virus (IBV) is a coronavirus that infects chickens, which exhibits a broad tropism for epithelial cells, infecting the tracheal mucosal epithelium, intestinal mucosal epithelium, and renal tubular epithelial cells. Utilizing single-cell RNA sequencing (scRNA-seq), we systematically examined cells in renal, bursal, and tracheal tissues following IBV infection and identified tissue-specific molecular markers expressed in distinct cell types. We evaluated the expression of viral RNA in diverse cellular populations and subsequently ascertained that distal tubules and collecting ducts within the kidney, bursal mucosal epithelial cells, and follicle-associated epithelial cells exhibit susceptibility to IBV infection through immunofluorescence. Furthermore, our findings revealed an upregulation in the transcription of proinflammatory cytokines IL18 and IL1B in renal macrophages as well as increased expression of apoptosis-related gene STAT in distal tubules and collecting duct cells upon IBV infection leading to renal damage. Cell-to-cell communication unveiled potential interactions between diverse cell types, as well as upregulated signaling pathways and key sender-receiver cell populations after IBV infection. Integrating single-cell data from all tissues, we applied weighted gene co-expression network analysis (WGCNA) to identify gene modules that are specifically expressed in different cell populations. Based on the WGCNA results, we identified seven immune-related gene modules and determined the differential expression pattern of module genes, as well as the hub genes within these modules. Our comprehensive data provides valuable insights into the pathogenesis of IBV as well as avian antiviral immunology.


Subject(s)
Cell Communication , Chickens , Coronavirus Infections , Gene Regulatory Networks , Infectious bronchitis virus , Single-Cell Analysis , Animals , Infectious bronchitis virus/genetics , Infectious bronchitis virus/physiology , Coronavirus Infections/virology , Coronavirus Infections/genetics , Poultry Diseases/virology , Poultry Diseases/genetics , Poultry Diseases/immunology , Sequence Analysis, RNA , Epithelial Cells/virology , Epithelial Cells/metabolism
11.
Virology ; 595: 110094, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38692133

ABSTRACT

Stress-induced immunosuppression (SIIS) is one of common problems in the intensive poultry industry, affecting the effect of vaccine immunization and leading to high incidences of diseases. In this study, the expression characteristics and regulatory mechanisms of miR-214 in the processes of SIIS and its influence on the immune response to avian influenza virus (AIV) vaccine in chicken were explored. The qRT-PCR results showed that serum circulating miR-214 was significantly differentially expressed (especially on 2, 5, and 28 days post immunization (dpi)) in the processes, so had the potential as a molecular marker. MiR-214 expressions from multiple tissues were closely associated with the changes in circulating miR-214 expression levels. MiR-214-PTEN regulatory network was a potential key regulatory mechanism for the heart, bursa of Fabricius, and glandular stomach to participate in the process of SIIS affecting AIV immune response. This study can provide references for further understanding of stress affecting immune response.


Subject(s)
Chickens , Influenza Vaccines , Influenza in Birds , MicroRNAs , PTEN Phosphohydrolase , Stress, Physiological , Animals , MicroRNAs/genetics , MicroRNAs/metabolism , Chickens/virology , Influenza Vaccines/immunology , Influenza in Birds/virology , Influenza in Birds/immunology , PTEN Phosphohydrolase/genetics , PTEN Phosphohydrolase/metabolism , Poultry Diseases/virology , Poultry Diseases/immunology , Immune Tolerance , Signal Transduction , Influenza A virus/immunology
12.
Vet Microbiol ; 293: 110094, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38636175

ABSTRACT

Infectious bursa disease (IBD) is an acute, highly contactable, lethal, immunosuppressive infectious disease caused by the Infectious bursa disease virus (IBDV). Currently, the emerged novel variant IBDV (nVarIBDV) and the sustainedly prevalent very virulent IBDV (vvIBDV) are the two most prevalent strains of IBDV in China. The antigenic properties of the two prevalent strains differed significantly, which led to the escape of nVarIBDV from the immune protection provided by the existing vvIBDV vaccine. However, the molecular basis of the nVarIBDV immune escape remains unclear. In this study, we demonstrated, for the first time, that residues 252, 254, and 256 in the PDE of VP2 are involved in the immune escape of the emerging nVarIBDV. Firstly, the IFA-mediated antigen-antibody affinity assay showed that PBC and PDE of VP2 could affect the affinity of vvIBDV antiserum to VP2, of which PDE was more significant. The key amino acids of PDE influencing the antigen-antibody affinity were also identified, with G254N being the most significant, followed by V252I and I256V. Then the mutated virus with point or combined mutations was rescued by reverse genetics. it was further demonstrated that mutations of V252I, G254N, and I256V in PDE could individually or collaboratively reduce antigen-antibody affinity and interfere with antiserum neutralization, with G254N being the most significant. This study revealed the reasons for the widespread prevalence of nVarIBDV in immunized chicken flocks and provided innovative ideas for designing novel vaccines that match the antigen of the epidemic strain.


Subject(s)
Birnaviridae Infections , Capsid Proteins , Chickens , Immune Evasion , Infectious bursal disease virus , Poultry Diseases , Infectious bursal disease virus/genetics , Infectious bursal disease virus/immunology , Animals , Chickens/virology , Capsid Proteins/genetics , Capsid Proteins/immunology , Poultry Diseases/virology , Poultry Diseases/immunology , Birnaviridae Infections/veterinary , Birnaviridae Infections/virology , Birnaviridae Infections/immunology , China , Antibodies, Viral/immunology , Mutation , Viral Vaccines/immunology , Viral Structural Proteins
13.
Poult Sci ; 103(6): 103741, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38670055

ABSTRACT

Fowl adenovirus serotype 4 (FAdV-4) infections result in substantial economic losses in the poultry industry. Recent findings have revealed that FAdV-4 significantly suppresses the host immune response upon infection; however, the specific viral and host factors contributing to this immunomodulatory activity remain poorly characterized. Moreover, diverse cell types exhibit differential immune responses to FAdV-4 infection. To elucidate cell-specific host responses, we performed transcriptomic analysis of FAdV-4 infected leghorn male hepatocellular (LMH) and chicken embryo fibroblast (CEF) cells. Although FAdV-4 replicated more efficiently in LMH cells, it provoked limited interferon-stimulated gene induction. In contrast, FAdV-4 infection triggered robust antiviral responses in CEF cells, including upregulation of cytosolic DNA sensing and interferon-stimulated genes. Knockdown of key cytosolic DNA sensing molecules enhanced FAdV-4 replication in LMH cells while reducing interferon-stimulated gene expression. Our findings reveal cell-specific virus-host interactions that provide insight into FAdV-4 pathogenesis while identifying factors that mediate antiviral immunity against FAdV-4.


Subject(s)
Adenoviridae Infections , Aviadenovirus , Chickens , Fibroblasts , Immunity, Innate , Poultry Diseases , Animals , Male , Fibroblasts/virology , Fibroblasts/immunology , Chick Embryo , Adenoviridae Infections/veterinary , Adenoviridae Infections/immunology , Adenoviridae Infections/virology , Poultry Diseases/virology , Poultry Diseases/immunology , Chickens/immunology , Aviadenovirus/physiology , Aviadenovirus/immunology , Serogroup , Hepatocytes/virology , Hepatocytes/immunology
14.
Poult Sci ; 103(6): 103640, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38688195

ABSTRACT

The transmission electron microscopy revealed a dendritic cell in the medulla of the chicken bursal follicle. This dendritic cell has a classical secretory machinery; therefore, it has been named a bursal secretory dendritic cell (BSDC). The corticomedullary epithelial arch (CMEA) encloses lymphoid-like cells, which can proliferate and after entering the medulla, begin to differentiate to immature, then mature BSDC, which discharges glycoprotein (gp). With the exhaustion of gp production, the BSDC rapidly transforms into a macrophage-like cell (Mal), which is an activated endocytic cell of innate immunity. The Mal drifts through the follicle-associated epithelium (FAE)-supporting cells into the FAE, and via FAE, the Mal is eliminated in the bursal lumen. The infectious bursal disease virus (IBDV) infection accelerates the maturation process of BSDC precursors, which results in acute emptying of CMEA and subsequently, numerous immature BSDC(s) emerge. The IBDV infection stops the gp discharge, and the gp appears in the virus-containing Mal. The Movat pentachrome staining recognizes the gp in the extracellular spaces of the medulla and after infection in the Mal. The BSDC is the primary target of the IBDV. During IBDV infection, a large number of suddenly formed Mal actively migrate into the cortex, initiating cytokine storm and recruiting heterophil granulocytes. During embryogenesis, the vimentin-positive, possibly embryonic dendritic cells provide a microenvironment for carbohydrate switch. Around hatching, these embryonic, temporary dendritic cells get the Fc receptor, which bind maternal IgY. The posthatched forms of BSDC(s) gradually replace the embryonic ones and bind their own IgY.


Subject(s)
Bursa of Fabricius , Chickens , Dendritic Cells , Infectious bursal disease virus , Animals , Bursa of Fabricius/virology , Dendritic Cells/physiology , Dendritic Cells/virology , Infectious bursal disease virus/physiology , Birnaviridae Infections/veterinary , Birnaviridae Infections/virology , Poultry Diseases/virology , Poultry Diseases/immunology
15.
Int J Biol Macromol ; 269(Pt 1): 131807, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38670189

ABSTRACT

Coccidiosis is an important parasitic protozoan disease in poultry farming, causing huge economic losses in the global poultry industry every year. MicroRNAs (miRNAs) are a class of RNA macromolecules that play important roles in the immune response to pathogens. However, the expression profiles and functions of miRNAs during Eimeria tenella (E. tenella) infection in chickens remain mostly uncharacterized. In this study, high-throughput sequencing of cecal tissues of control (JC), resistant (JR), and susceptible (JS) chickens led to the identification of 35 differentially expressed miRNAs among the three groups. Functional enrichment analysis showed that the differentially expressed miRNAs were mainly associated with the TGF-beta, NF-kB, and Jak-STAT signaling pathways. Notably, gga-miR-2954 was found to be significantly upregulated after coccidial infection. Functional analysis showed that gga-miR-2954 inhibited the production of the inflammatory cytokines IL-6, IL-1ß, TNF-α, and IL-8 in sporozoite-stimulated DF-1 cells. Mechanistically, we found that gga-miR-2954 targeted the RORC gene and that RORC promoted the inflammatory response in sporozoite-stimulated DF-1 cells. In conclusion, our study was the first to identify differentially expressed miRNAs in chicken cecal tissue during E. tenella infection and found that gga-miR-2954 regulates the host immune response to coccidial infection in chickens by targeting the RORC gene.


Subject(s)
Chickens , Coccidiosis , Eimeria tenella , Gene Expression Profiling , MicroRNAs , Poultry Diseases , Animals , Cecum/parasitology , Cell Line , Coccidiosis/veterinary , Coccidiosis/immunology , Coccidiosis/genetics , Coccidiosis/parasitology , Cytokines/metabolism , Cytokines/genetics , Gene Expression Regulation , Inflammation/genetics , Inflammation/immunology , Inflammation/parasitology , MicroRNAs/genetics , Poultry Diseases/parasitology , Poultry Diseases/genetics , Poultry Diseases/immunology , Signal Transduction , Transcriptome , Male , Female
16.
Viruses ; 16(4)2024 04 07.
Article in English | MEDLINE | ID: mdl-38675911

ABSTRACT

Zika virus (ZIKV) remains a public health concern, with epidemics in endemic regions and sporadic outbreaks in new areas posing significant threats. Several mosquito-borne flaviviruses that can cause human illness, including West Nile, Usutu, and St. Louis encephalitis, have associations with birds. However, the susceptibility of chickens to ZIKV and their role in viral epidemiology is not currently known. We investigated the susceptibility of chickens to experimental ZIKV infection using chickens ranging from 1-day-old chicks to 6-week-old birds. ZIKV caused no clinical signs in chickens of all age groups tested. Viral RNA was detected in the blood and tissues during the first 5 days post-inoculation in 1-day and 4-day-old chicks inoculated with a high viral dose, but ZIKV was undetectable in 6-week-old birds at all timepoints. Minimal antibody responses were observed in 6-week-old birds, and while present in younger chicks, they waned by 28 days post-infection. Innate immune responses varied significantly between age groups. Robust type I interferon and inflammasome responses were measured in older chickens, while limited innate immune activation was observed in younger chicks. Signal transducer and activator of transcription 2 (STAT2) is a major driver of host restriction to ZIKV, and chicken STAT2 is distinct from human STAT2, potentially contributing to the observed resistance to ZIKV infection. The rapid clearance of the virus in older chickens coincided with an effective innate immune response, highlighting age-dependent susceptibility. Our study indicates that chickens are not susceptible to productive ZIKV infection and are unlikely to play a role in the ZIKV epidemiology.


Subject(s)
Chickens , Immunity, Innate , Poultry Diseases , Zika Virus Infection , Zika Virus , Animals , Chickens/virology , Zika Virus Infection/immunology , Zika Virus Infection/virology , Zika Virus/immunology , Disease Susceptibility , Poultry Diseases/virology , Poultry Diseases/immunology , Age Factors , Antibodies, Viral/blood , RNA, Viral/genetics
17.
Viruses ; 16(4)2024 04 14.
Article in English | MEDLINE | ID: mdl-38675946

ABSTRACT

Infectious bronchitis virus (IBV) is a highly contagious Gammacoronavirus causing moderate to severe respiratory infection in chickens. Understanding the initial antiviral response in the respiratory mucosa is crucial for controlling viral spread. We aimed to characterize the impact of IBV Delmarva (DMV)/1639 and IBV Massachusetts (Mass) 41 at the primary site of infection, namely, in chicken tracheal epithelial cells (cTECs) in vitro and the trachea in vivo. We hypothesized that some elements of the induced antiviral responses are distinct in both infection models. We inoculated cTECs and infected young specific pathogen-free (SPF) chickens with IBV DMV/1639 or IBV Mass41, along with mock-inoculated controls, and studied the transcriptome using RNA-sequencing (RNA-seq) at 3 and 18 h post-infection (hpi) for cTECs and at 4 and 11 days post-infection (dpi) in the trachea. We showed that IBV DMV/1639 and IBV Mass41 replicate in cTECs in vitro and the trachea in vivo, inducing host mRNA expression profiles that are strain- and time-dependent. We demonstrated the different gene expression patterns between in vitro and in vivo tracheal IBV infection. Ultimately, characterizing host-pathogen interactions with various IBV strains reveals potential mechanisms for inducing and modulating the immune response during IBV infection in the chicken trachea.


Subject(s)
Chickens , Coronavirus Infections , Gene Expression Profiling , Infectious bronchitis virus , Poultry Diseases , Trachea , Animals , Trachea/virology , Trachea/immunology , Chickens/virology , Infectious bronchitis virus/physiology , Infectious bronchitis virus/immunology , Coronavirus Infections/veterinary , Coronavirus Infections/immunology , Coronavirus Infections/virology , Poultry Diseases/virology , Poultry Diseases/immunology , Poultry Diseases/genetics , Epithelial Cells/virology , Epithelial Cells/immunology , Transcriptome , Host-Pathogen Interactions/immunology , Host-Pathogen Interactions/genetics , Virus Replication , Specific Pathogen-Free Organisms
18.
Vaccine ; 42(15): 3410-3419, 2024 May 31.
Article in English | MEDLINE | ID: mdl-38641498

ABSTRACT

The application of recombinant herpesvirus of turkey, expressing the H9 hemagglutinin gene from low pathogenic avian influenza virus (LPAIV) H9N2 and the avian orthoavulavirus-1 (AOAV-1) (commonly known as Newcastle Disease virus (NDV)) fusion protein (F) as an rHVT-H9-F vaccine, is an alternative to currently used classical vaccines. This study investigated H9- and ND-specific humoral and mucosal responses, H9-specific cell-mediated immunity, and protection conferred by the rHVT-H9-F vaccine in specific pathogen-free (SPF) chickens. Vaccination elicited systemic NDV F- and AIV H9-specific antibody response but also local antibodies in eye wash fluid and oropharyngeal swabs. The ex vivo H9-specific stimulation of splenic and pulmonary T cells in the vaccinated group demonstrated the ability of vaccination to induce systemic and local cellular responses. The clinical protection against a challenge using a LPAIV H9N2 strain of the G1 lineage isolated in Morocco in 2016 was associated with a shorter duration of shedding along with reduced viral genome load in the upper respiratory tract and reduced cloacal shedding compared to unvaccinated controls.


Subject(s)
Antibodies, Viral , Chickens , Influenza A Virus, H9N2 Subtype , Influenza Vaccines , Influenza in Birds , Virus Shedding , Animals , Influenza A Virus, H9N2 Subtype/immunology , Influenza A Virus, H9N2 Subtype/genetics , Chickens/immunology , Influenza in Birds/prevention & control , Influenza in Birds/immunology , Influenza Vaccines/immunology , Influenza Vaccines/administration & dosage , Antibodies, Viral/immunology , Antibodies, Viral/blood , Virus Shedding/immunology , Specific Pathogen-Free Organisms , Newcastle disease virus/immunology , Newcastle disease virus/genetics , Poultry Diseases/prevention & control , Poultry Diseases/immunology , Poultry Diseases/virology , Immunity, Cellular , Herpesvirus 1, Meleagrid/immunology , Herpesvirus 1, Meleagrid/genetics , Vaccination/methods , Immunity, Humoral , Genetic Vectors/immunology , Immunogenicity, Vaccine , Vaccines, Synthetic/immunology , Vaccines, Synthetic/administration & dosage , Hemagglutinin Glycoproteins, Influenza Virus/immunology , Hemagglutinin Glycoproteins, Influenza Virus/genetics
19.
Avian Dis ; 68(1): 10-17, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38687102

ABSTRACT

The relationship between passive immunity and the development of false layer syndrome (FLS) and its associated lesions was investigated in this study by comparing the long-term reproductive effects of an infectious bronchitis virus (IBV) DMV/1639 wild-type strain and the GA08 vaccine in birds with and without maternal antibodies. There was a clear protective effect provided by maternal antibodies against both the early vaccination and challenge. It was also observed that vaccination at an early age, in the absence of maternal antibodies, can induce reproductive issues, such as reduced egg production and FLS-associated lesions (e.g., cystic oviduct and egg yolk coelomitis). This might indicate that maternal antibodies and the timing of IBV infection are more important in the generation of FLS than the IBV strain type.


Mitigación del síndrome de la falsa ponedora mediante anticuerpos maternos contra el virus de la bronquitis infecciosa. En este estudio se investigó la relación entre la inmunidad pasiva y el desarrollo del síndrome de la falsa ponedora (FLS) y sus lesiones asociadas comparando los efectos reproductivos a largo plazo de una cepa de tipo silvestre DMV/1639 del virus de la bronquitis infecciosa (IBV) y la cepa vacunal GA08, en aves con y sin anticuerpos maternos. Hubo un claro efecto protector proporcionado por los anticuerpos maternos tanto contra la vacunación temprana como contra el desafío. También se observó que la vacunación a una edad temprana, en ausencia de anticuerpos maternos, puede inducir problemas reproductivos, como una reducción de la producción de huevo y lesiones asociadas al síndrome de la falsa ponedora (p. ej., oviducto quístico y celomitis de yema de huevo). Esto podría indicar que los anticuerpos maternos y el momento de la infección por el virus de la bronquitis infecciosa son más importantes en la generación del síndrome de la falsa ponedora que el tipo de cepa del virus de la bronquitis infecciosa.


Subject(s)
Antibodies, Viral , Chickens , Coronavirus Infections , Infectious bronchitis virus , Poultry Diseases , Infectious bronchitis virus/immunology , Animals , Poultry Diseases/virology , Poultry Diseases/immunology , Female , Coronavirus Infections/veterinary , Coronavirus Infections/virology , Coronavirus Infections/immunology , Immunity, Maternally-Acquired , Viral Vaccines/immunology , Viral Vaccines/administration & dosage
20.
PLoS One ; 19(4): e0302555, 2024.
Article in English | MEDLINE | ID: mdl-38683795

ABSTRACT

Clostridial dermatitis (CD), caused by Clostridium septicum, is an emerging disease of increasing economic importance in turkeys. Currently, there are no effective vaccines for CD control. Here, two non-toxic domains of C. septicum alpha toxin, namely ntATX-D1 and ntATX-D2, were identified, cloned, and expressed in Escherichia coli as recombinant subunit proteins to investigate their use as potential vaccine candidates. Experimental groups consisted of a Negative control (NCx) that did not receive C. septicum challenge, while the adjuvant-only Positive control (PCx), ntATX-D1 immunization (D1) and ntATX-D2 immunization (D2) groups received C. septicum challenge. Turkeys were immunized subcutaneously with 100 µg of protein at 7, 8 and 9 weeks of age along with an oil-in-water nano-emulsion adjuvant, followed by C. septicum challenge at 11 weeks of age. Results showed that while 46.2% of birds in the PCx group died post-challenge, the rate of mortality in D1- or D2-immunization groups was 13.3%. The gross and histopathological lesions in the skin, muscle and spleen showed that the disease severity was highest in PCx group, while the D2-immunized birds had significantly lower lesion scores when compared to PCx. Gene expression analysis revealed that PCx birds had significantly higher expression of pro-inflammatory cytokine genes in the skin, muscle and spleen than the NCx group, while the D2 group had significantly lower expression of these genes compared to PCx. Peripheral blood cellular analysis showed increased frequencies of activated CD4+ and/or CD8+ cells in the D1 and D2-immunized groups. Additionally, the immunized turkeys developed antigen-specific serum IgY antibodies. Collectively, these findings indicate that ntATX proteins, specifically the ntATX-D2 can be a promising vaccine candidate for protecting turkeys against CD and that the protection mechanisms may include downregulation of C. septicum-induced inflammation and increased CD4+ and CD8+ cellular activation.


Subject(s)
Bacterial Toxins , Clostridium Infections , Clostridium septicum , Dermatitis , Poultry Diseases , Recombinant Proteins , Turkeys , Animals , Turkeys/immunology , Clostridium septicum/immunology , Clostridium Infections/prevention & control , Clostridium Infections/immunology , Clostridium Infections/veterinary , Poultry Diseases/prevention & control , Poultry Diseases/immunology , Poultry Diseases/microbiology , Bacterial Toxins/immunology , Recombinant Proteins/immunology , Recombinant Proteins/administration & dosage , Dermatitis/prevention & control , Dermatitis/immunology , Dermatitis/veterinary , Bacterial Vaccines/immunology , Bacterial Vaccines/administration & dosage , Immunization
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