interleukin-8 and Poultry-Diseases

Marek's disease is a contagious lymphoma of chickens caused by Marek's disease virus (MDV). MDV replicates in chicken lymphocytes and establishes latency within and transforms chicken CD4+ T-cells. Transformed T-cells are seen as skin leukosis or as lymphomas in visceral organs. A major focus of our laboratory is the functional study of genes flanking the origin of replication. This origin (OriLyt) is contained within the repeats flanking the unique long (UL) region of the genome (IRL and TRL). To the left of this Ori are genes associated with MDV latent/transforming infection [1.8-kb RNA family, pp14, Meq), and to the right (UL) are genes associated with early stages of MDV lytic infection [BamHI-H-encoded protein (Hep), pp38/pp24, Mys]. During latency, MDV suppresses lytic gene expression and has evolved mechanisms for blocking the apoptosis of latently-infected CD4+ T-cells. Of the genes expressed during MDV latency and in the transformed cell, the Meq (Marek's EcoRI-Q-encoded protein) has been shown to block apoptosis and transactivate gene expression. Upon reactivation to lytic infection, we have found that splice variants of Meq predominate and that these forms lack several of the domains important to Meq trans-activation and trans-repression. We have found that rightward from the origin of replication, a family genes, including phosphoprotein 38 (pp38) are expressed during early stages of reactivation. Three separate open reading frames (Hep, Mys, and pp38) are encoded by distinct transcripts from this region. We are now determining the kinetics of expression of these transcripts and their relative abundance during reactivation.

Topics: Animals; Apoptosis; CD4-Positive T-Lymphocytes; Gene Expression Regulation; Interleukin-8; Kinetics; Mardivirus; Marek Disease; Phosphoproteins; Poultry; Poultry Diseases; Transcription, Genetic; Virus Replication

Salmonella enterica (SE) is a major foodborne bacterial pathogen in the United States, commonly found as the normal flora of various animals that is attributed to causing at least 1.2 million infections annually. Poultry plays a major role in disseminating SE through direct contact with live animals and consumption of contaminated products. Vaccinating poultry against SE is a sustainable approach that can reduce SE in the host, preventing future infections in humans. An intracellular autolytic SE serovar Typhimurium vaccine (STLT2

Topics: Animals; Chickens; Humans; Interleukin-12; Interleukin-6; Interleukin-8; Poultry Diseases; RNA, Ribosomal, 16S; Salmonella enterica; Salmonella Infections, Animal; Salmonella typhimurium; Salmonella Vaccines

Wooden breast (WB) myopathy in broiler chickens is a growing challenge for the poultry industry. Previous multi-omic data have implied that the pathogenesis of WB is associated with the activation of immune system and inflammatory response. However, the intricate mechanisms are not fully understood. This study was therefore conducted to systematically investigate the morphology, expression of cytokines as well as the underlying signaling pathways regulating the inflammatory response in pectoralis major (PM) muscle of WB myopathic broilers.. wHistopathological changes, increased plasma creatine kinase and lactate dehydrogenase activities, elevated myeloperoxidase activity and overproduction of nitric oxide in muscle indicated the enhancement of muscle damage and inflammation in WB broilers. The messenger RNA (mRNA) expressions of inflammatory cytokines were dysregulated in PM muscle and contents of interleukin (IL)-1β, IL-8 and tumor necrosis factor-α were increased in serum of WB myopathic broilers, indicating this myopathy was associated with immune disorder and systemic inflammation response. Additionally, toll-like receptor (TLR) levels were upregulated, the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathway was activated and the mRNA expression levels of downstream inflammatory mediators were enhanced in PM muscle of WB myopathy affected birds.. The results indicated an immune disorder and a systemic inflammation response in WB myopathic broilers, which might be related to a synergetic effect of TLRs and NF-κB pathway. © 2020 Society of Chemical Industry.

Topics: Animals; Chickens; Cytokines; Interleukin-8; Male; Muscular Diseases; NF-kappa B; Poultry Diseases; Tumor Necrosis Factor-alpha; Up-Regulation

IL-6, IL-8, and C-C motif chemokine ligand 2 (CCLi2) are important factors in inflammatory and immune responses. To investigate their relationships in the spleen and cecum and between coccidiosis-infected and uninfected states, we performed quantitative real-time PCR to compare the relative expression difference of IL-6, IL-8, and CCLi2 in the same tissues between the infection and control groups. In addition, the correlations of the relative expression levels of these 3 genes were determined in the same and different tissues within the same group. The results showed that the expression levels of IL-6, IL-8, and CCLi2 in the spleen and cecum of the infected group were all higher than those of the uninfected group (P < 0.05). The correlation coefficients among the IL-6, IL-8, and CCLi2 expression levels in the spleen or cecum were all positive in both the infection and control groups. In the spleen tissues, CCLi2 expression was strongly correlated with IL-6 and IL-8 in the uninfected group (P < 0.01), and the correlation coefficients reached 0.853 (R

Topics: Animals; Avian Proteins; Cecum; Chemokine CCL2; Coccidiosis; Eimeria tenella; Gene Expression; Interleukin-6; Interleukin-8; Ligands; Poultry Diseases; Spleen

Avian pathogenic Escherichia coli (APEC) is an important pathogen that causes avian colibacillosis in poultry. APEC infection can lead to pathological changes in chicken trachea. The type VI secretion system (T6SS) of APEC contribute to the pathogenicity of APEC. However, whether T6SS plays a role in infection of the trachea remains unclear. We constructed mutant strain Δhcp2a by the Red recombination method system. The role of hcp2a (the structural secretion components and secretory protein of the T6SS) in the infection of trachea was investigated. The mutation strain displayed a significant increase in biofilm formation and a decrease in resistance to chicken serum. Moreover, RNA sequencing analyses showed that infection of chicken tracheal epithelium by the mutant strain Δhcp2a induced differential expression of genes. The result also showed that 14 genes (13 genes were downregulated) were enriched in cytokine-cytokine receptor interaction signalling pathway at 12 and 24 h post infection. The mutation Δhcp2a resulted in significant decreases in the bacterial loads in trachea at 6 and 12 h post infection. Real-time PCR analyses showed that the hcp2a mutation downregulated the expression of IL8 and IL1β at mRNA level in chicken tracheal epithelium. Our results indicate that mutation of hcp2a influenced genes expression of the cytokine-cytokine receptor interaction pathway by decreasing APEC colonization in the trachea.

Topics: Animals; Bacterial Load; Chickens; Epithelium; Escherichia coli; Escherichia coli Infections; Escherichia coli Proteins; Gene Expression Regulation; Interleukin-1beta; Interleukin-8; Mutation; Poultry Diseases; Trachea; Type VI Secretion Systems; Virulence Factors

Avian salmonellosis caused by

Topics: Animals; Bacterial Proteins; Beclin-1; Caco-2 Cells; Chickens; Cytokines; HeLa Cells; Host-Pathogen Interactions; Humans; Interleukin-8; MAP Kinase Signaling System; NF-kappa B; Poultry Diseases; Salmonella enterica; Salmonella enteritidis; Salmonella Infections, Animal; Serogroup; Transfection; Tumor Necrosis Factor-alpha

The chicken upper respiratory tract is the portal of entry for respiratory pathogens including avian influenza virus (AIV). There is a paucity of information about the role of airway epithelial cells in the induction of antiviral responses in the chicken trachea. A better understanding of the role of these cells in the initiation of innate responses may improve prophylactic or therapeutic strategies for control of viral infections. The present study aimed to characterize antiviral innate responses in chicken tracheal epithelial cells (cTECs) induced by TLR ligands. The results demonstrated that stimulation of cTECs with TLR ligands induced antiviral responses, and subsequently reduced the replication of AIV in cTECs. Additionally, stimulated cTECs were able to influence the function of other cells such as macrophages. Overall, these results provided evidence that cTECs mount antiviral responses after stimulation with TLR ligands through IRF7 and NF-κB signaling pathways, leading to activation of other cells, such as macrophages.

Topics: Animals; Avian Proteins; Chickens; Epithelial Cells; Immunity, Innate; Influenza A virus; Influenza in Birds; Interferon Regulatory Factors; Interleukin-1beta; Interleukin-8; Ligands; Macrophages; Poly I-C; Poultry Diseases; Toll-Like Receptor 3; Toll-Like Receptor 4; Trachea

Infectious laryngotracheitis virus (ILTV) is an alphaherpesvirus that infects chickens, causing upper respiratory tract disease and significant losses to poultry industries worldwide. Glycoprotein G (gG) is a broad-range viral chemokine-binding protein conserved among most alphaherpesviruses, including ILTV. A number of studies comparing the immunological parameters between infection with gG-expressing and gG-deficient ILTV strains have demonstrated that expression of gG is associated with increased virulence, modification of the amount and the composition of the inflammatory response, and modulation of the immune responses toward antibody production and away from cell-mediated immune responses. The aims of the current study were to examine the establishment of infection and inflammation by ILTV and determine how gG influences that response to infection.

Topics: Animals; Antibodies, Viral; Chemokines; Chickens; Cytokines; Herpesviridae Infections; Herpesvirus 1, Gallid; Inflammation Mediators; Interleukin-8; Organ Culture Techniques; Poultry Diseases; Protein Binding; Specific Pathogen-Free Organisms; Trachea; Viral Envelope Proteins; Virulence

The avian coronavirus causes infectious bronchitis (IB), which is one of the most serious diseases affecting the avian industry worldwide. However, there are no effective strategies for controlling the IB virus (IBV) at present. Therefore, development of novel antiviral treatment strategies is urgently required. As reported, astragalus polysaccharides (APS) have potential antiviral effects against several viruses; however, the antiviral effect of APS against IBV remains unclear. In this study, we explored whether APS had the potential to inhibit IBV infectionby utilizing several in vitro experimental approaches. To this end, the effect of APS on the replication of IBV was examined in chicken embryo kidney (CEK) cells. Viral titers were calculated by using the plaque formation assay, and the cytotoxicity of APS was tested by utilizing a Cell Counting Kit-8 assay. The expression of viral mRNA and cytokine (IL-1β, IL-6, IL-8 and TNF-α) mRNA transcripts was determined by real-time quantitative RT-PCR(qRT-PCR). IBV titers in infected CEK cells treated with APS were significantly reduced in a dose-dependent manner, indicating that APS inhibited IBV replication in vitro. We also found that the decreased viral replication after APS treatment was associated with reduced mRNA levels of the cytokines IL-1B, IL-6, IL-8 and TNF-α. In conclusion, these results suggest that APS exhibit antiviral activities against IBV and it may represent a potential therapeutic agent for inhibiting the replication of IBV.

Topics: Animals; Antiviral Agents; Astragalus Plant; Cell Line; Cell Proliferation; Chickens; Coronavirus Infections; Cytokines; Infectious bronchitis virus; Interleukin-1beta; Interleukin-6; Interleukin-8; Plant Extracts; Polysaccharides; Poultry Diseases; RNA, Messenger; Tumor Necrosis Factor-alpha; Viral Load; Viral Plaque Assay; Virus Replication

Hydropericardium syndrome and inclusion body hepatitis, together called hydropericardium-hepatitis syndrome, are acute infectious diseases found in chickens. These diseases are caused primarily by fowl adenovirus serotype 4 (FAdV-4) strains. In this study, we isolated a FAdV-4 strain (SD0828) from clinically diseased chickens and phylogenetically analyzed the L1 loops of the hexon protein sequences in 3-week-old specific pathogen-free chickens and ducks infected intramuscularly and orally, determining differences in the pathogenicity by observing clinical signs and gross and histological lesions. We also detected the viral load in tissue samples. Postinfection necropsy showed that all chickens but no ducks exhibited typical necropsy lesions. Additionally, all chickens infected intramuscularly died within 2 days postinfection (dpi), and all those infected orally died within 5 dpi, whereas no infected ducks died before 28 dpi. Quantitative real-time polymerase chain reaction analysis was used to determine the viral load in the tissues of hearts, livers, spleens, lungs, and kidneys and in cloacal cotton swabs from infected chickens and ducks at 1, 2, 3, 5, 7, 14, 21, and 28 dpi. The greatest number of viral DNA copies was found in the livers of infected chickens, yet no virus was found in any samples from infected ducks. In addition, the viral load increased over time in both chicken and duck embryo fibroblasts (CEFs and DEFs, respectively); in the former, replication speed was significantly greater than in the latter. Innate immune responses were also studied, both

Topics: Adenoviridae Infections; Animals; Aviadenovirus; Chickens; DEAD Box Protein 58; DNA, Viral; Ducks; Immunity, Innate; Interleukin-6; Interleukin-8; Poultry Diseases; Viral Load; Virus Replication

Both CCR5 and CXCR4 are important chemokine receptors and take vital role in migration, development and distribution of T cells, however, whether they will influence the process of T cell infiltration into bursa of Fabricius during infectious bursal disease virus (IBDV) infection is unclear. In the current study, CCR5 and CXCR4 antagonists, Maraviroc and AMD3100, were administrated into chickens inoculated with IBDV, and the gene levels of IBDV VP2, CCR5, CXCR4 and related cytokines were determined by real-time PCR. The results showed that large number of T cells began to migrate into the bursae on Day 3 post infection with IBDV and the mRNA of chemokine receptors CCR5 and CXCR4 began to increase on Day 1. Moreover, antagonist treatments have increased the VP2, CCR5 and CXCR4 gene transcriptions and influenced on the gene levels of IL-2, IL-6, IL-8, IFN-γ, TGF-β4, MHC-I and MDA5. In conclusion, the chemokine receptors CCR5 and CXCR4 might influence virus replication during IBDV infection and further study would focus on the interaction between chemokine receptors and their ligands.

Topics: Animals; Benzylamines; Bursa of Fabricius; CCR5 Receptor Antagonists; Cell Movement; Chickens; Cyclams; Cyclohexanes; Cytokines; Heterocyclic Compounds; Infectious bursal disease virus; Interferon-gamma; Interleukin-2; Interleukin-6; Interleukin-8; Maraviroc; Poultry Diseases; Real-Time Polymerase Chain Reaction; Receptors, CCR5; Receptors, Chemokine; Receptors, CXCR4; RNA, Messenger; T-Lymphocytes; Transcription, Genetic; Transforming Growth Factor beta; Triazoles; Viral Structural Proteins; Virus Replication

Toll-like receptor 3 (TLR3) is one of the TLRs whose ligand is double-stranded RNA (dsRNA). Infectious bursal disease virus (IBDV) is a dsRNA virus that could be recognized by TLR3. The purpose of this study was to determine the role of the virulence of IBDV on the expression of chicken TLR3 (chTLR3). For this purpose, the levels of chTLR3 expression and its downstream effectors, Interferon β (IFN-β) and Interleukin 8 (IL-8), were detected and analyzed after infection of IBDV field isolates with differential virulence in vitro (chicken embryo fibroblast and/or chicken peripheral blood mononuclear cells) and in vivo (commercial Three-Yellow chicken). The results showed that chTLR3 was activated by IBDV, resulting in the expression of antiviral IFN-β and chemokine IL-8. The expression of chTLR3, IFN-β, and IL-8 correlated well with the virulence of IBDV as the more virulent the IBDV strain that was used, the more pronounced was the expression of chTLR3, IFN-β, and IL-8. These results suggest that chTLR3 is involved in the pathogenesis of IBDV in commercial chickens and its downstream effectors (IFN-β and IL-8) might play an important role in this process.

Topics: Animals; Birnaviridae Infections; Cells, Cultured; Chickens; Gene Expression Regulation; Infectious bursal disease virus; Interferon-beta; Interleukin-8; Poultry Diseases; RNA, Messenger; RNA, Viral; Time Factors; Toll-Like Receptor 3; Virulence; Virus Replication

Vaccination has been widely used to reduce the Salmonella burden in poultry and subsequently the transmission to humans. Concerning turkey, there is little knowledge on the immune response to colonization and invasion by Salmonella species or about efficacy of vaccination and involved immune mechanisms. In the present study, turkeys were vaccinated at the day of hatch and infected with Salmonella Typhimurium (ST) or Enteritidis (SE) field strains three weeks later. A control group was kept uninfected. After challenge infection, bacterial counts in the cecal content, liver and spleen were determined 7 and 14days post infection. They were often statistically significantly lower in vaccinated poults than in non-vaccinated ones. Production of iNOS, and the cytokines IL-8, IL-10 and IFN-γ were reduced in vaccinated birds. However, neither the influx of CD4+, CD8α+ and CD28+ cells into cecal mucosa after infection nor the antibody response were statistically significantly altered in vaccinated birds.

Topics: Animals; Cecum; Cytokines; Interleukin-10; Interleukin-8; Liver; Poultry Diseases; Salmonella enteritidis; Salmonella Infections, Animal; Salmonella typhimurium; Salmonella Vaccines; Spleen; Turkeys; Vaccination

Astragalus polysaccharides (APS) are biological macromolecules extracted from Astragalus species that have strong immunoregulatory properties. In this study, APS were employed as an adjuvant for an avian infectious bronchitis virus (IBV) vaccine, and its effects on the cellular immune and humoral immune responses to vaccination in chicken were investigated. One hundred and fifty chicken were randomly divided into five groups (n = 30, each group). The chickens in all groups, except for the unvaccinated control group, were vaccinated with an IBV DNA vaccine. Three of the four vaccinated groups were administered different doses of APS (APSL, 10 mg/kg; APSM, 50 mg/kg; and APSH, 100 mg/kg) after the first vaccination, and the remaining vaccinated group served as a control, without any additional treatment. At 14, 28, and 42 days after the first vaccination, serum anti-IBV antibody titers; peripheral lymphocyte proliferation; and the mRNA expression of IL-1β, IL-2, IL-8, and TNF-α in the spleen were assessed by enzyme-linked immunosorbent assay (ELISA), the cell counting kit-8 (CCK-8), and real time quantitative RT-PCR (qRT-PCR), respectively. At most time points, the titer of IBV-specific antibodies, lymphocyte proliferation, and IL-1β, IL-2, IL-8, and TNF-α mRNA expression levels were higher in three APS groups than in the vaccine control group, and these increases were dose-dependent. These data suggest that APS could be used as an adjuvant for IBV vaccination to provide better protection against IBV infection.

Topics: Adaptive Immunity; Adjuvants, Immunologic; Animals; Antibodies, Viral; Astragalus Plant; Cell Proliferation; Chickens; Coronavirus Infections; Cytokines; Enzyme-Linked Immunosorbent Assay; Infectious bronchitis virus; Interleukin-1beta; Interleukin-2; Interleukin-8; Lymphocytes; Peptide Fragments; Plant Extracts; Polysaccharides; Poultry Diseases; RNA, Messenger; Spleen; Time Factors; Tumor Necrosis Factor-alpha; Vaccination; Vaccines, DNA; Viral Vaccines

Interleukins are reported to be valuable immunostimulants in enhancing the immune efficiency of conventional vaccines. In this study, the effect of expression of interleukin 8 (IL-8) on the immune response of ducks to avian influenza vaccine was investigated. The results showed that the serum antibody titer, lymphocyte transformation efficiency and serum interferon gamma (IFN-γ) level of ducks injected with avian influenza vaccine along with a plasmid expressing duck IL-8 were higher than those of ducks injected with conventional immunostimulant Astragalus polysaccharide (APS) or empty plasmid. Therefore, the duck IL-8 may be used as a good immunostimulant to enhance the immune efficiency of avian influenza vaccine in ducks.

Topics: Animals; Antibodies, Viral; Ducks; Influenza A Virus, H5N1 Subtype; Influenza in Birds; Influenza Vaccines; Interleukin-8; Poultry Diseases

Bacterial artificial chromosome (BAC) vectors were first developed to facilitate propagation and manipulation of large DNA fragments. This technology was later used to clone full-length genomes of large DNA viruses to study viral gene function. Marek's disease virus (MDV) is a highly oncogenic herpesvirus that causes rapid induction of T-cell lymphomas in chickens. Based on the virus's ability to cause disease in vaccinated chickens, MDV strains are classified into pathotypes, with the most virulent strains belonging to the very virulent plus (vv+) pathotype. Here we report the construction of BAC clones of 686 (686-BAC), a vv+ strain of MDV. Transfection of DNA isolated from two independent clones into duck embryo fibroblasts resulted in recovery of infectious virus. Pathogenesis studies showed that the BAC-derived 686 viruses were more virulent than Md5, a vv strain of MDV. With the use of a two-step red-mediated mutagenesis process, both copies of viral interleukin 8 (vIL-8) were deleted from the MDV genome, showing that 686-BACs were amenable to mutagenesis techniques. The generation of BAC clones from a vv+ strain of MDV is a significant step toward understanding molecular basis of MDV pathogenesis.

Topics: Animals; Cells, Cultured; Chickens; Chromosomes, Artificial, Bacterial; DNA, Viral; Ducks; Escherichia coli; Gene Deletion; Herpesvirus 2, Gallid; Interleukin-8; Marek Disease; Mutagenesis; Polymerase Chain Reaction; Poultry Diseases; Transfection

Our previous studies have illustrated three strains of duck-origin H5N1 highly pathogenic avian influenza viruses (HPAIVs) had varying levels of pathogenicity in ducks (Sun et al., 2011). However, the host immune response of ducks infected with those of H5N1 HPAIVs was unclear. Here, we compared viral distribution and mRNA expression of immune-related genes in ducks following infection with the two HPAIV (A/Duck/Guangdong/212/2004, DK212 and A/Duck/Guangdong/383/2008, DK383). DK383 could replicate in the tested tissue of ducks (brain, spleen, lungs, cloacal bursa, kidney, and pancreas) more rapid and efficiently than DK212 at 1 and 2 days post-inoculation. Quantitative real-time PCR analysis showed that the expression levels of TLR3, IL-6, IL-8, and MHC class II in brains were higher than those of respective genes in lungs during the early stage of post infection. Furthermore, the expression levels of IL-6 and IL-8 in the brain of ducks following infection with DK383 were remarkably higher than those of ducks infected with DK212, respectively. Our results suggest that the shift in the H5N1 HPAIVs to increased virulence in ducks may be associated with efficient and rapid replication of the virus, accompanied by early destruction of host immune responses. These data are helpful to understand the underlying mechanism of the different outcome of H5N1 HPAIVs infection in ducks.

Topics: Animals; Ducks; Influenza A Virus, H5N1 Subtype; Influenza in Birds; Interleukin-6; Interleukin-8; Lung; Poultry Diseases; Real-Time Polymerase Chain Reaction; Spleen; Virulence

Genetic line and diet affect chicken heterophil activity and gene expression, and the combination of these factors can enhance disease resistance. This study evaluated the effects of immune modulating diets on heterophil/lymphocyte (H/L) ratio and heterophil chemokine expression in distinct genetic lines. Fayoumi and Leghorn chickens were fed a basal diet or immune modulating diets enhanced with β-glucans, ascorbic acid, or corticosterone. H/L ratios and heterophil gene expression in response to in vitro stimulation with Salmonella enteritidis (SE) were evaluated on days 1, 3, 7, and 21 of diet treatment. The stress-mimicking corticosterone diet influenced H/L ratio in the Leghorn line, but not the Fayoumi line, suggesting resistance to stress-induced immunosuppression in the Fayoumi line. Leghorn line H/L ratios were increased on days 1 and 3 of corticosterone diet treatment, but not days 7 or 21. Expression of CXCLi2 by SE stimulated heterophils was higher in the Leghorn line, suggesting that Leghorns rely more heavily on inflammatory response than do Fayoumis. Corticosterone diet was associated with reduced CXCLi2 expression in heterophils from both lines. Dietary β-glucan or ascorbic acid did not affect H/L ratio or CXCLi2 expression, suggesting that benefits of these immunomodulators may not be evident in healthy birds.

Topics: Animals; Ascorbic Acid; Avian Proteins; beta-Glucans; Chickens; Corticosterone; Diet; Gene Expression; Granulocytes; Immunologic Factors; Interleukin-8; Poultry Diseases; Salmonella enteritidis; Salmonella Infections, Animal; Species Specificity

In commercial poultry production, there is a lack of natural flora providers since chickens are hatched in the clean environment of a hatchery. Events occurring soon after hatching are therefore of particular importance, and that is why we were interested in the development of the gut microbial community, the immune response to natural microbial colonization, and the response to Salmonella enterica serovar Enteritidis infection as a function of chicken age. The complexity of chicken gut microbiota gradually increased from day 1 to day 19 of life and consisted of Proteobacteria and Firmicutes. For the first 3 days of life, chicken cecum was protected by increased expression of chicken β-defensins (i.e., gallinacins 1, 2, 4, and 6), expression of which dropped from day 4 of life. On the other hand, a transient increase in interleukin-8 (IL-8) and IL-17 expression could be observed in chicken cecum on day 4 of life, indicating physiological inflammation and maturation of the gut immune system. In agreement, the response of chickens infected with S. Enteritidis on days 1, 4, and 16 of life shifted from Th1 (characterized mainly by induction of gamma interferon [IFN-γ] and inducible nitric oxide synthase [iNOS]), observed in younger chickens, to Th17, observed in 16-day-old chickens (characterized mainly by IL-17 induction). Active modification of chicken gut microbiota in the future may accelerate or potentiate the maturation of the gut immune system and increase its resistance to infection with different pathogens.

Topics: Aging; Animals; beta-Defensins; Cecum; Chickens; Cytokines; Enteritis; Enzyme-Linked Immunosorbent Assay; Gastrointestinal Tract; Immunity, Innate; Interleukin-17; Interleukin-8; Polymerase Chain Reaction; Poultry Diseases; Proteobacteria; RNA, Ribosomal, 16S; Salmonella enteritidis; Salmonella Infections, Animal; Th1 Cells; Th17 Cells

To characterize 12 Salmonella Virchow isolates from human and avian sources to begin to determine the genetic relationships within the serovar, determine its capacity to invade and induce inflammatory responses in human intestinal epithelial cells and investigate its ability to colonize the chicken gastrointestinal tract.. Multi-Locus Sequence Typing (MLST) revealed that 11 isolates belonged to sequence type 16 (ST16). Pulsed Field Gel Electrophoresis (PFGE) grouped the isolates into two main clusters. All isolates contained genes associated with virulence determined through PCR virulotyping. All the S. Virchow isolates had the ability to invade human epithelial cells and elicit high levels of production of the pro-inflammatory chemokine interleukin-8 (IL-8). Experimental infection of poultry showed S. Virchow colonizes the caeca and spleen.. Isolates within the serovar show high levels of genetic relatedness regardless of the source. The data indicates S. Virchow is an invasive and inflammatory serovar, consistent with its association with invasive salmonellosis in humans.. The poultry infection experiment included in this study shows S. Virchow can colonize the gastrointestinal tract rapidly and to high levels with the chickens showing no clinical signs of infection. The asymptomatic colonization of chickens indicates an increased ability of S. Virchow to enter the food chain undetected and cause human salmonellosis which because of the invasive and inflammatory nature of S. Virchow seen during the Caco2 invasion assay and previous studies showing its invasive nature in humans and increasing resistance to antibiotics is a public health concern.

Topics: Animals; Bacterial Typing Techniques; Caco-2 Cells; Chickens; Chlorocebus aethiops; Electrophoresis, Gel, Pulsed-Field; England; Epithelial Cells; Humans; Interleukin-8; Multilocus Sequence Typing; Poultry; Poultry Diseases; Public Health; Salmonella enterica; Salmonella Infections; Vero Cells; Virulence

Campylobacter (C.) jejuni and C. coli can cause gastrointestinal disorders in humans characterized by acute inflammation. Inflammatory signals are initiated during interaction between these pathogens and human intestinal cells, but nothing is known about the stimulation of avian intestinal cells by Campylobacter. Interleukin-8 (IL-8) as a proinflammatory chemokine plays an important role in mobilizing cellular defence mechanism. IL-8 mRNA expression in both human intestinal cells (INT 407) and primary intestinal chick cells (PIC) was determined by quantitative real-time RT-PCR. The secretion of IL-8 protein by INT407 was measured using ELISA. Although C. jejuni and C. coli are considered to be harmless commensals in the gut of birds, the avian Campylobacter isolates investigated were able to induce the proinflammatory IL-8 in PIC as well as in INT407. In an in vitro system, C. jejuni as well as C. coli were able to induce IL-8 mRNA in PIC. Relation between the virulence properties like toxin production, the ability to invade and to survive in Caco-2 cells and the level of IL-8 mRNA produced by INT 407 and PIC after infection with Campylobacter strains was also investigated.

Topics: Animals; Campylobacter; Campylobacter coli; Campylobacter Infections; Campylobacter jejuni; Cell Line; Chickens; DNA, Complementary; Enzyme-Linked Immunosorbent Assay; Humans; Immunohistochemistry; Interleukin-8; Intestinal Mucosa; Poultry Diseases; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Specific Pathogen-Free Organisms; Virulence

Campylobacter jejuni is one of the leading causes of food-borne gastroenteritis. Because of the high prevalence of C. jejuni in poultry, poultry meat is considered a major source of C. jejuni infections for humans. However, it is not known whether all poultry-associated C. jejuni strains are capable of causing disease in humans. Four different virulence properties of C. jejuni strains were compared between 20 poultry isolates and 24 human isolates. Strains were chosen based on their PFGE pattern to represent a heterogeneous population. The isolates were compared for their ability to invade and induce interleukin-8 (IL-8) production in T84 cells, their production of functional cytolethal distending toxin (CDT) using HEp-2 cells, and their sodium deoxycholate resistance. All four virulence factors were present among strains of human and poultry origin, with strong differences observed among strains. For invasion and IL-8 induction, no difference was observed between the two populations. However, on average, human isolates arrested more HEp-2 cells in their cell cycle than did the poultry isolates (P=0.041), suggesting higher CDT production by the former. The ability to survive 16 000 mug sodium deoxycholate ml(-1) was significantly more pronounced (P=0.006) among human isolates than poultry isolates, although all strains possessed the cmeABC operon. These data suggest that all four virulence properties are widespread among C. jejuni isolates, but that a higher degree of bile-salt resistance and more pronounced CDT production are associated with strains causing enteritis in humans.

Topics: Animals; Anti-Bacterial Agents; Bacterial Toxins; Campylobacter Infections; Campylobacter jejuni; Cell Line, Tumor; Chickens; Deoxycholic Acid; Drug Resistance, Bacterial; Genes, Bacterial; Humans; Interleukin-8; Operon; Poultry; Poultry Diseases; Virulence

Marek's disease virus (MDV) is an alpha-herpesvirus that causes rapid development of T-cell lymphomas in chickens. MDV-encoded vIL-8 is homologous to the cellular IL-8 gene, and its function in MDV pathogenesis has yet to be determined. Using overlapping cosmid clone-based technology, we have generated an MDV vIL-8 deletion mutant virus, rMd5/delta vIL-8. In vivo experiments with this mutant virus demonstrated that deletion of vIL-8 results in attenuation of the virus and induction of significantly less gross tumor, both in viscera and nerves, when compared to the parental virus. Reintroduction of the vIL-8 gene in the genome of the mutant virus restored the virulence of the virus to the wild-type levels, indicating that vIL-8 plays a role in MDV-induced pathogenesis. In this study, we show that there is a significant difference in the reduction of B cells and activation of T cells in the spleen cells of chickens inoculated with parental rMd5 and vIL-8 deletion mutant virus. These results indicate that vIL-8 is involved in the early phase of pathogenesis, presumably by attracting target cells to the initial site of infection. In addition, protection studies with the vIL-8 mutant virus showed that this mildly virulent virus protects susceptible maternal antibody-positive viruses at a higher level than the commonly used serotype 1 CVI988 vaccine. These results confirm the potential of partially attenuated viruses as vaccines against very virulent plus strains and the usefulness of recombinant DNA technology to generate the next generation of MDV vaccines.

Topics: Animals; Antibodies, Monoclonal; Chickens; Flow Cytometry; Gene Deletion; Herpesvirus 2, Gallid; Immunity, Cellular; Immunohistochemistry; Interleukin-8; Marek Disease; Poultry Diseases; Virulence

Marek's disease, a lymphoproliferative disease of chickens, is caused by an alphaherpesvirus, Marek's disease virus (MDV). This virus encodes a virokine, vIL-8, with general homology to cellular CXC chemokines such as interleukin-8 (IL-8) and Gro-alpha. To study the function of vIL-8 gene, we deleted both copies of vIL-8 residing in the terminal repeat long and internal repeat long region of the viral genome and generated a mutant virus with vIL-8 deleted, rMd5/DeltavIL-8. Growth kinetics study showed that vIL-8 gene is dispensable for virus replication in cell culture. In vivo, the vIL-8 gene is involved in early cytolytic infections in lymphoid organs, as evidenced by limited viral antigen expression of rMd5/DeltavIL-8. However, the rMd5/DeltavIL-8 virus is unimpaired in virus replication in the feather follicle epithelium. vIL-8 does not appear to be important for establishment of latency, since rMd5/DeltavIL-8 and the wild-type virus have similar viremia titers at 14 days postinfection, a period when the virus titer comes primarily from reactivated latent genomes. Nevertheless, because of the impaired cytolytic infections, the overall transformation efficiency of the virus with vIL-8 deleted is much lower, as reflected by the reduced number of transformed cells at 5 weeks postinoculation and the presence of fewer gross tumors. Importantly, the revertant virus that restored the expression of vIL-8 gene also restored the wild-type phenotype, indicating the deficient phenotypes are results of vIL-8 deletion. One of the interesting differences between the MDV vIL-8 gene and its cellular counterpart is the presence of a DKR (Asp-Lys-Arg) motif instead of ELR (Glu-Leu-Arg) preceding the invariable CXC motif. To study the significance of this variation, we generated recombinant MDV, rMd5/vIL-8-ELR, carrying the ELR motif. Both in vitro and in vivo studies revealed that the DKR motif is as competent as ELR in pathogenesis of MDV.

Topics: Animals; Chickens; Female; Gene Deletion; Herpesvirus 2, Gallid; Interleukin-8; Lymphoid Tissue; Male; Marek Disease; Poultry Diseases; Recombination, Genetic; Viral Proteins; Virus Latency; Virus Replication

We recently showed that increased in vitro heterophil functional efficiency translates to increased in vivo resistance to a systemic Salmonella enteritidis (SE) infection utilizing a parental pair of broiler chickens (lines A and B) and the F1 reciprocal crosses (C and D). Heterophils produce cytokines and modulate acute protection against Salmonella in young poultry. Therefore, we hypothesize that heterophils from SE-resistant chickens (A and D) have the ability to produce an up-regulated pro-inflammatory cytokine response compared to that of heterophils from SE-susceptible chickens (B and C). In this study, heterophils were isolated from day-old chickens and treated with either RPMI-1640 (as the control), or phagocytic agonists (SE, or SE opsonized with either normal chicken serum or immune serum against SE) and cytokine mRNA expression assessed using real-time quantitative reverse transcription-polymerase chain reaction. Heterophils from SE-resistant chickens (A and D) had significantly higher levels of pro-inflammatory cytokine (interleukin (IL)-6, IL-8, and IL-18) mRNA expression upon treatment with all agonists compared to heterophils from SE-susceptible lines (B and C). Further, heterophils from SE-resistant chickens had significantly decreased mRNA expression levels of transforming growth factor-beta4, an anti-inflammatory cytokine, when compared to heterophils from SE-susceptible chickens. These data indicate cytokine gene expression in heterophils may be a useful parameter in determining resistance to Salmonella, as indicated by our previous in vivo SE studies. Therefore, heterophil functional efficiency and cytokine production may be useful biomarkers for poultry breeders to consider when developing new immunocompetent lines of birds.

Topics: Animals; Chickens; Cytokines; Disease Susceptibility; Gene Expression; Immunocompetence; Interleukin-18; Interleukin-6; Interleukin-8; Neutrophils; Poultry Diseases; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Salmonella Infections, Animal; Transforming Growth Factor beta

Marek's disease (MD) in chickens is caused by MD herpesvirus (MDV), which induces T cell lymphomas. The early pathogenesis of MDV infection is characterized by a primary infection in B lymphocytes followed by infection of activated T lymphocytes. It has been speculated that a MDV-encoded homologue of interleukin-8 (vIL-8) may be important to attract activated T lymphocytes to infected B lymphocytes. Recently, more virulent strains of MDV have emerged, named very virulent plus (vv+)MDV, that cause earlier and more prolonged cytolytic infections compared to less virulent strains. In this report, it was found that vIL-8 mRNA expression in vivo was increased in very virulent (vv) and vv+MDV strains compared to mild (m) and virulent (v) strains, and could not be detected in two attenuated MDV strains examined using very sensitive real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assays. In order to identify potential mechanisms for the increased vIL-8 mRNA expression in more virulent strains, and lack thereof in attenuated strains, the vIL-8 gene and putative promoter sequences upstream of the vIL-8 gene were compared from 10 different MDV strains, including attenuated derivatives. Only the JM-16 strain (both non-attenuated and attenuated) and attenuated 584A (584Ap80C) encoded a predicted vIL-8 gene sequence different from all other strains examined. Within the putative vIL-8 gene promoter sequence, there was little difference among the non-attenuated strains; however significant deletions were identified in the attenuated JM-16/p71, Md11 (R2/23), and 584Ap80C strains. Additionally, these deletions were located within a previously hypothetical open reading frame (ORF) named LORF4. Rapid amplification of cDNA ends identified a full-length transcript of LORF4 in the MDV-transformed lymphoblastoid cell line MSB-1, and deletions within this ORF caused truncated predicted proteins in 4 out of 6 attenuated MDV strains examined.

Topics: Amino Acid Sequence; Animals; Base Sequence; Cell Line; Chickens; Deoxyribonuclease BamHI; Gene Deletion; Herpesvirus 2, Gallid; Interleukin-8; Marek Disease; Molecular Sequence Data; Open Reading Frames; Poultry Diseases; Sequence Analysis, DNA; Transcription, Genetic; Virulence

Increased resistance to Salmonella enteritidis (SE) organ infectivity in chickens can be conferred by the prophylactic administration of SE-immune lymphokines (ILK). Resistance is associated with an enhanced heterophilic accumulation within 4 h of ILK injection. In these studies, the role of IL-8 in ILK-mediated heterophil recruitment during SE infections in young chickens was investigated. Heterophil accumulation was enhanced 2-4 h after the i.p. injection of both ILK and SE (ILK/SE) when compared to the control chicks. An i.p. injection of a rabbit polyclonal anti-human IL-8 antibody significantly (P < 0.01) reduced the accumulation of heterophils in the peritoneum after the injection of ILK/SE. Injections of preimmune rabbit IgG had no effect on peritoneal heterophil numbers. Within 2 h of injection of ILK/SE, a ten-fold increase in heterophil chemotactic activity was found in the peritoneal lavage fluid from these chicks compared to the saline control chicks. Pretreatment, with the anti-IL-8 antibody, of the peritoneal lavage fluids collected from the ILK/SE-treated chicks dramatically reduced this heterophil chemotactic activity. Treatment of the lavage fluids from all groups with preimmune IgG had no effect on heterophil chemotaxis. Additionally, pretreatment of ILK with the anti-human IL-8 antibody had no effect on heterophil chemotaxis. The results from these experiments suggest that IL-8 is produced locally by the host in response to both the SE infection and the ILK. With these studies, it was established that IL-8 is a major chemotactic factor produced by the host, which aids in mediating the ILK/SE-induced recruitment of heterophils to the site of SE invasion.

Topics: Animals; Chemotaxis, Leukocyte; Chickens; Granulocytes; Inflammation; Interleukin-8; Lymphokines; Neutrophils; Peritoneal Lavage; Peritoneum; Poultry Diseases; Salmonella enteritidis; Salmonella Infections, Animal; Spleen; T-Lymphocytes; Time Factors

Eight chicken cytokine genes (IL-1beta, IL-2, IL-8, IL-15, IFN-alpha, IFN-gamma, TGF-beta4, lymphotactin) were evaluated for their adjuvant effect on a suboptimal dose of an Eimeria DNA vaccine carrying the 3-1E parasite gene (pcDNA3-1E). Chickens were given two subcutaneous injections with 50 microg of the pcDNA3-1E vaccine plus a cytokine expression plasmid 2 weeks apart and challenged with Eimeria acervulina 1 week later. IFN-alpha (1 microg) or 10 microg of lymphotactin expressing plasmids, when given simultaneously with the pcDNA3-1E vaccine, significantly protected against body weight loss induced by E. acervulina. Parasite replication was significantly reduced in chickens given the pcDNA3-1E vaccine along with 10 microg of the IL-8, lymphotactin, IFN-gamma, IL-15, TGF-beta4, or IL-1beta plasmids compared with chickens given the pcDNA3-1E vaccine alone. Flow cytometric analysis of duodenum intraepithelial lymphocytes showed chickens that received the pcDNA3-1E vaccine simultaneously with the IL-8 or IL-15 genes had significantly increased CD3+ cells compared with vaccination using pcDNA3-1E alone or in combination with the other cytokine genes tested. These results indicate that the type and the dose of cytokine genes injected into chickens influence the quality of the local immune response to DNA vaccination against coccidiosis.

Topics: Adjuvants, Immunologic; Animals; Chickens; Coccidiosis; Drug Evaluation, Preclinical; Duodenum; Eimeria; Genetic Vectors; Interferon-alpha; Interferon-gamma; Interferons; Interleukin-1; Interleukin-15; Interleukin-2; Interleukin-8; Interleukins; Lymphokines; Parasite Egg Count; Poultry Diseases; Sialoglycoproteins; Specific Pathogen-Free Organisms; Transforming Growth Factor beta; Vaccination; Vaccines, DNA; Weight Gain