Chicken Peripheral Blood Mononuclear Cells Research Articles

THE IN VITRO AND IN VIVO CYTOTOXICITY OF LIPOSOME-ENCAPSULATED DICHLOROMETHYLENE-DIPHOSPHONATE ON CHICKEN MACROPHAGES AND THE POTENTIAL OF USING MACROPHAGE-DEPLETED CHICKENS AS A MODEL FOR REOVIRUS INFECTION

The liposome-encapsulated dichloromethylene-diphosphonate ([Formula: see text]) is known to be toxic to the cultured macrophages and has selective cytotoxic effects on macrophages in rats and mice, but has minimal adverse effects on the non-phagocytic cells following various administration routes. In this study, the cytotoxicity of [Formula: see text] on chicken macrophages was investigated. Similar to findings related to other mammals, the [Formula: see text] was toxic to the macrophages originating from the chicken peripheral blood mononuclear cells as indicated by the MTT (3-(4, 5-dimethylthiazole-2-yl) 2, 5-diphonyltetrazolium bromide) cleavage assays. After administered with [Formula: see text], chicken spleens were rapidly and progressively reduced in sizes. This reduction was more severely gross than with the experimental course. One day post-treatment, the histopathology showed that most of the macrophages around the peri-ellipsoid white pulp had undergone marked cellular necrosis and lysis, and more severe lesions appeared with the loss of lymphoid and reticuloendothelial tissues at 3 and 5 days post-treatment. White pulp which has been replaced by red pulp in the spleen were noticed at 3 and 5 days post-treatment. The flow cytometric assays further confirmed the depletion of the macrophages in the chicken spleens following the administration of [Formula: see text], when compared with those of the controls. Furthermore, the replication of the avian reovirus (ARV) in the spleens was significantly reduced in the macrophage-depleted chickens administered with [Formula: see text]. Thus, depletion of the macrophages with [Formula: see text] followed by functional assesses in the macrophage-depleted chicken could be used as a model to confirm the involvement of the macrophages in ARV replication.

In-vitro assessment of differential cytokine gene expression in response to infections with Egyptian classic and variant strains of highly pathogenic H5N1 avian influenza virus

In Egypt, two distinct genetic groups of HPAI H5N1 viruses are co-circulating: classic 2.2.1/C sub-clade and antigenic drift variant 2.2.1.1 clade isolated from vaccinated poultry flocks. The response of chicken innate immunity to both genotypes is not investigated, so far. In this study, expression of immune related genes (IL1b, IL4, IL6, IL8, IL10, IL18, IFNα and IFNγ) after infecting chicken macrophage cell line (HD11) and chicken peripheral blood Mononuclear cells (PBMC) with a classic and a variant strains was assayed using quantitative reverse-transcription real-time polymerase chain reaction assays (qRT-PCR). In HD11, the variant strain induced higher levels of IL1b and IL8 at 6hours post infection (hpi), IL4 at 24 / 48hpi and IFNα at 48hpi than the classic strain. Conversely, the classic strain induced about 10-fold increase of IFNγ at 24 and 48hpi and the virus replicated at higher level than the variant strain. The results of PBMC infection were similar to that reported from HD11 except for IFNγ gene expression that was higher at variant strain infected cells than that infected with the classic strain. After 24hpi skewing the innate immune response toward anti-inflammatory (humoral-associated) cytokines was different between HD11 (through IL4) and PBMC (through IL10). To sum up, the classic strain produced less cytokines which may indicate adaptation to evade the recognition by the innate immune system and explain its higher pathogenicity.

Expression of cytokines in chicken peripheral blood mononuclear cells after stimulation by probiotic bacteria and Campylobacter jejuni in vitro

The aim of this study was to delineate the ability of selected probiotic strains to reduce invasion by Campylobacter jejuni in peripheral blood mononuclear cells (PMBCs). PMBCs were isolated from chicken blood and exposed in vitro to probiotic strains Enterococcus faecium EF55, E. faecium AL41, E. faecium H31, Lactobacillus fermentum AD1, and infected with C. jejuni CCM6191. The mRNA expression levels for selected cytokines were determined at 24 and 48 h postinfection (pi) using reverse transcriptase-polymerase chain reaction. The level of expression of IL-1β, LITAF, and MIP1-β was upregulated (P < 0.05; P < 0.01; P < 0.001) in LAD + CJ (L. fermentum AD1 + C. jejuni) group when compared to control (C), CJ (C. jejuni), and other combined groups (EF55 + CJ, AL 41 + CJ, H31 + CJ) 24 h pi. Overall, L. fermentum AD1 and E. faecium AL41 demonstrated the highest immunomodulatory activity against campylobacter infection in vitro.

Synergy of lipopolysaccharide and resiquimod on type I interferon, pro-inflammatory cytokine, Th1 and Th2 response in chicken peripheral blood mononuclear cells

Toll-like receptors (TLRs) recognize conserved molecular structures of invading pathogens and initiate an immune response to curtail the infection prior to the development of more powerful and specific adaptive immunity. Understanding the interactions between different TLRs in terms of immune response genes is a pre-requisite for using various TLR agonists alone or in combination as adjuvants or as stand-alone agents against various diseases. Lipopolysaccharide (LPS) and resiquimod (R-848) are TLR agonists that are recognized by TLR4 and TLR7, respectively. In this study, the effect of LPS and/or R-848 on chicken peripheral blood mononuclear cells (PBMCs) was investigated. LPS and R-848 synergistically up-regulated the transcripts of interferon-β (IFN-β), IFN-γ, IL-4 and IL-1β as compared to the individual response (P<0.05). The results indicate that these agonists synergistically interact and enhance type-I IFN, pro-inflammatory cytokine as well as Th1 and Th2 responses in chicken PBMCs, suggesting their potential as an adjuvant candidate to be used in combination with various poultry vaccines.

Isolation and characterization of peripheral blood-derived endothelial progenitor cells from broiler chickens

Peripheral blood-derived endothelial progenitor cells (EPCs) have been extensively studied in mammals but the isolation and characterization of EPCs in avian species have not been reported. In this study, chicken peripheral blood mononuclear cells (PBMNCs) were cultured under conditions favoring endothelial-specific differentiation for 2 weeks. One heterogeneous cell population dominated by spindle-shaped cells (early EPCs) and one homogeneous cell population exhibiting cobblestone-like morphology (endothelial outgrowth cells, EOCs) appeared sequentially.Quantitative polymerase chain reaction (PCR) showed the expression of several progenitor and endothelial cell markers such as CD133, VEGFR-2 and CD31 in both cell populations. However, CD34, another progenitor marker, was undetectable in either freshly isolated PBMNCs or cultured cells. The endothelial phenotype of the EOCs was further identified by acetylated low-density lipoprotein/lectin double staining, and in vitro tube formation. Collectively, these data demonstrate that chicken EPCs can be isolated and cultured from PBMNCs and suggest that EPCs obtained from peripheral blood may originate mainly from the CD34− subpopulation.

Both host and parasite MIF molecules bind to chicken macrophages via CD74 surface receptor

Macrophage migration inhibitory factor (MIF) is recognized as a soluble protein that inhibits the random migration of macrophages and plays a pivotal immunoregulatory function in innate and adaptive immunity. Our group has identified both chicken and Eimeria MIFs, and characterized their function in enhancing innate immune responses during inflammation. In this study, we report that chicken CD74 (ChCD74), a type II transmembrane protein, functions as a macrophage surface receptor that binds to MIF molecules. First, to examine the binding of MIF to chicken monocytes/macrophages, fresh isolated chicken peripheral blood mononuclear cells (PBMCs) were stimulated with rChIFN-γ and then incubated with recombinant chicken MIF (rChMIF). Immunofluorescence staining with anti-ChMIF followed by flow cytometry revealed the binding of MIF to stimulated PBMCs. To verify that ChCD74 acts as a surface receptor for MIF molecules, full-length ChCD74p41 was cloned, expressed and its recombinant protein (rChCD74p41) transiently over-expressed with green fluorescent protein in chicken fibroblast DF-1 cells. Fluorescence analysis revealed a higher population of cells double positive for CD74p41 and rChMIF, indicating the binding of rChMIF to DF-1 cells via rChCD74p41. Using a similar approach, it was found that Eimeria MIF (EMIF), which is secreted by Eimeria sp. during infection, bound to chicken macrophages via ChCD74p41 as a surface receptor. Together, this study provides conclusive evidence that both host and parasite MIF molecules bind to chicken macrophages via the surface receptor ChCD74.

Cloning and Characterization of a 7 Transmembrane Receptor from the Adherent Cells of Chicken Peripheral Blood Mononuclear Cells

A cDNA encoding a 7 transmembrane (7TM) receptor gene from the adherent cells of chicken peripheral blood mononuclear cells (PBMC) was cloned and characterized. The open reading frame of the chicken-7TM (Ch-7TM) receptor gene was 1008 nucleotides long, encoding a protein of 335 amino acid residues with a molecular mass of approximately 37.1 kDa. Hydrophobic stretches indicated the presence of 7 TM domains. Moreover, the complete nucleotide sequences encoding 7TM of duck (Du-7TM) and goose (Go-7TM), corresponding to the open reading frame of Ch-7TM, were determined. Each of the Du- and Go-7TM encoding regions comprised 990 nucleotides, representing an 18-nucleotide deletion in alignment with the Ch-7TM encoding region, resulting in a 6-amino-acid deletion at the 3′-end. No signal peptides were predicted. Six phosphorylation sites were predicted and conserved for all three 7TMs. The proteins of the three 7TMs were similar, with 11 conserved cysteine residues. No glycosylation sites could be predicted. The results of the pairwise comparisons indicated that the Ch-7TM encoding region and Ch-7TM protein were the least similar to those of Du- and Go-7TMs. These results were in accordance with those of the phylogenetic analysis, which indicated that the Du- and Go-7TM encoding regions clustered, but were separated from the Ch-7TM encoding region. Monoclonal antibody B28D5 was prepared from spleens of mice immunized with the bacterially expressed N-terminal (55 amino acid residues) region of the Ch-7TM protein for further use. Double staining with B28D5 and KUL01 suggested that Ch-7TM was expressed in subsets of the adherent cells, among which a subset that was recognized with both antibodies was likely of monocyte and macrophage lineage. However, the fluorescence intensities of B28D5 and, particularly, KUL01 decreased after the adherent cells were incubated for additional 48 h.

Proteomic analysis of chicken peripheral blood mononuclear cells after infection by Newcastle disease virus.

Characteristic clinical manifestations of Newcastle disease include leukopenia and immunosuppression. Peripheral blood mononuclear cells (PBMCs) are the main targets of Newcastle disease virus (NDV) infection. To survey changes in proteomic expression in chicken PBMCs following NDV infection, PBMC proteins from 30 chickens were separated using two-dimensional electrophoresis (2-DE) and subjected to mass spectrometry analysis. Quantitative intensity analysis showed that the expression of 78 proteins increased more than two-fold. Thirty-five proteins exhibited consistent changes in expression and 13 were identified as unique proteins by matrix assisted laser desorption ionization-time of flight mass spectrometer/mass spectrometer including three that were down-regulated and 10 that were up-regulated. These proteins were sorted into five groups based on function: macromolecular biosynthesis, cytoskeleton organization, metabolism, stress responses, and signal transduction. Furthermore, Western blot analysis confirmed the down-regulation of integrin-linked kinase expression and up-regulation of lamin A production. These data provide insight into the in vivo response of target cells to NDV infection at the molecular level. Additionally, results from this study have helped elucidate the molecular pathogenesis of NDV and may facilitate the development of new antiviral therapies as well as innovative diagnostic methods.

Recombinant flagellin and its cross-talk with lipopolysaccharide – Effect on pooled chicken peripheral blood mononuclear cells

Toll-like receptors (TLRs) are one of the types of pattern recognition receptors (PRRs) that recognize conserved pathogen molecules. TLRs link innate and adaptive arms of immune system and are implicated in the development of defense against invading pathogens. Lipopolysaccharide (LPS) and flagellin are recognized by TLR4 and TLR5, respectively. In this study, the effect of flagellin and lipopolysaccharide alone and in combination on chicken peripheral blood mononuclear cells (PBMCs) was investigated. The FliC gene of Salmonella typhimurium was expressed in a prokaryotic expression system and the recombinant flagellin was used to stimulate the chicken PBMCs. A combination of recombinant flagellin and LPS synergistically upregulated nitric oxide production, IL-12 and IL-6 expression but antagonistically down regulated IL-4 expression in comparison to recombinant flagellin alone. The results indicate that these agonists synergistically interact and enhance macrophage function and promote Th1 immune response in chicken PBMCs.

Analysis of immune-related gene expression in chicken peripheral blood mononuclear cells following Salmonella enterica serovar Enteritidis infection in vitro

We examined mRNA expression of eight genes, TLR4, TLR5, TLR15, interleukin (IL)-1β, IL-6, transforming growth factor-β4 (TGF-β4), CXCLi2, and a macrophage inflammatory protein (MIP) family chemokine called CCLi2, in peripheral blood mononuclear cells (PBMCs) isolated from the blood of chickens after in vitro exposure to Salmonella enterica serovar Enteritidis (SE). The chickens of four Chinese native lines, Qingjiaoma, Sanhuang, Wugu, and Xueshanma, were evaluated for mRNA expression levels at 2 and 4h post-infection using quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). TLR4 and TLR15 mRNA were in particular highly expressed in PBMCs of Wugu and Xueshanma chickens exposed to SE, while TLR5 was expressed less in the Sanhuang chickens than in others. Breed effect was significant (P<0.05) for IL-1β, IL-6, CXCLi2, and CCLi2 mRNA expression, all of which were expressed to a greater extent in Wugu and Xueshanma than in the other two lines. These findings demonstrate the difference of mRNA expression profiles for innate immune molecules in PBMCs infected to SE among different lines.

In vitro propagation of chicken anemia virus in chicken peripheral blood mononuclear cells

chicken anemia virus; propagation; mononuclear cells; PCR.

Enhanced Egress of Intracellular Eimeria tenella Sporozoites by Splenic Lymphocytes from Coccidian-Infected Chickens

Egress, which describes the mechanism that some intracellular parasites use to exit from parasitophorous vacuoles and host cells, plays a very important role in the parasite life cycle and is central to Eimeria propagation and pathogenesis. Despite the importance of egress in the intracellular parasite's life cycle, very little information is known on this process compared to other steps, e.g., invasion. The present study was conducted to investigate the interplay between the host adaptive immune system and Eimeria egression. Splenic lymphocytes or soluble immune factors were incubated with parasite-infected host cells for 3 or 5 h, and the percentage of egress was calculated according to an established formula. Viability of egressed parasites and host cells was tested using trypan blue exclusion and annexin V and propidium iodide staining, respectively. We found that premature egression of sporozoites from Eimeria tenella-infected primary chicken kidney cells or from chicken peripheral blood mononuclear cells occurred when the cells were cocultured in vitro with spleen lymphocytes from E. tenella-infected chickens but not when they were cocultured with splenocytes from uninfected chickens. Eimeria-specific antibodies and cytokines (gamma interferon [IFN-γ], interleukin-2 [IL-2], and IL-15), derived from E. tenella-primed B and T lymphocytes, respectively, were capable of promoting premature egress of sporozoites from infected host cells. Both egressed parasites and host cells were viable, although the latter showed reduced reinvasion ability. These results suggest a novel, immune-mediated mechanism that the host exploits to interrupt the normal Eimeria life cycle in vivo and thereby block the release of mature parasites into the environment.

Flavonoid, Phenol and Polysaccharide Contents of Echinacea Purpurea L. and Its Immunostimulant Capacity In Vitro

Echinacea purpurea L. (EP) is a plant originally used by native Americans to treat respiratory infections and have long been used to aid in wound healing and to enhance the immune system. The purpose of this study was to demonstrate the ability of EP extracts to stimulate the production of nitric oxide (NO) and TNF-α as well as to evaluate the cell viability by the use of chicken peripheral blood mononuclear cells (PBMCs) and RAW 264.7 macrophages in vitro. The polysaccharides content of EP was 162.2±8.4 mg/g dry weight (DW)The EP extracted with a 55% ethanol at 55℃ contained 22.3 ±1.0 mg gallic acid equivalent/g DW of total phenolic compounds and 86.0 ± 4.6 mg quercetin equivalent/g of flavonoid content. The result of cell viability assay showed that 89% and 81% of chicken PBMCs and RAW 264.7 macrophages, respectively, were survived under the treatment of 3.2 mg/ml of EP extracts by microculture tetrazolium assays (MTAs). Moreover, the cell inhibitory activity did not show the 50% cyto-toxicity effect (IC50). In addition, NO production by stimulated chicken PBMCs and RAW 264.7 macrophages exhibited individually the linear relations with the concentration of EP extracts, and they were approximately 70 and 60 μM in 800 μg/ml, respectively. The tendency with the release of TNF-α by RAW 264.7 macrophages also corresponded with EP extracts concentration. It is concluded, EP extracts act as an immunostimulant for both tested cells but have no serious effect on inhibiting chicken PBMCs viability.

The Immune Enhancement of Sodium Lauryl Sulfoacetate in Chickens

The purpose of this study is to investigate feasibility of sodium lauryl sulfoacetate (SLS) as an immunoadjuvant in chickens. After treating with 62.5, 125, 250, or 500 μg/mL SLS in vitro, lymphocyte proliferation assay of chicken peripheral blood mononuclear cells showed that the OD570 values of all experimental groups, as well as Con A-stimulated group, were significantly higher than that of the untreated control group. After injection with 1.0, 2.0, or 4.0 mg/kg of SLS for 3 consecutive days, chickens were vaccinated with an attenuated vaccine against Newcastle disease virus (NDV), and the immunoadjuvant effects of SLS were evaluated on the basis of immune organ index, antibody response, and CD4 +/CD8 + T-cell ratio. The results confirmed that SLS could enhance NDV-specific antibody response and increase CD4 +/CD8 + T-cell ratio in vivo. Furthermore, SLS could improve NDV-specific antibody response in thiamphenicol-treated chickens. These data indicate that SLS not only can improve humoral immune response but also reverse the immunosuppressive effect of thiamphenicol in chickens.

Immune response of peripheral blood mononuclear cells to avian pathogenicEscherichia coli

The exposure of chicken peripheral blood mononuclear cells (PBMCs) to pathogen can induce a rapid change in both pro-inflammatory and anti-inflammatory cytokine gene expression. Chicken PBMCs were infected with avian pathogenicEscherichia coli (APEC) and the isogenicfimH mutants of APEC, PBMCs immune response was determined by semi-quantitative RT-PCR. The results demonstrated that PBMCs were expressed TLR1/6/10, TLR3, TLR4, TLR5 and TLR7. APEC and non pathogenicE. Coli strains were significantly (P<0.05) increased the expression of IL-1β, IL-8, IL-18 and TGF-β4 cytokines compared to the isogenicfimH mutants. In addition, the opsonized avian pathogenicE. Coli strains were significantly (P<0.05) increased the expression of cytokines compared to the non-opsonized strains, whereas the opsonized avian non-pathogenicE. Coli strain was significantly (P < 0.05) decreased the expression of cytokines compared to the non-opsonized strain. PBMCs are important in APEC infection, the increase in cytokines expression may play important role in the regulation of immune response.

Phosphorothioate oligodeoxyribonucleotides induce in vitro proliferation of chicken B-cells

The study aimed to evaluate short synthetic oligodeoxyribonucleotides (ODN) as inducers of proliferation of chicken peripheral blood mononuclear cells (PBMC) and to identify the proliferating cells. A panel of different ODN; with phosphodiester and/or phosphorothioate backbone, with and without CpG-motifs, was therefore assessed for in vitro induction of proliferation. Six complete phosphorothioate ODN induced proliferation of PBMC while the complete phosphodiester or chimeric phosphodiester/phosphorohiate ODN did not. Moreover, CpG-motifs were not essential for induction of proliferation as responses to CpG-ODN were similar to those of their GpC controls. Two stimulatory phosphorothioate ODN were also used in phosphodiester form. In this comparison, only the phosphorothioate ODN were active despite the identical nucleotide sequences of their phosphodiester counterparts. In order to deliver DNA to the cytoplasm and decrease degradation of ODN by nucleases, stimulating as well as inactive ODN were treated with lipofectin prior to induction. However, proliferative responses were not influenced by lipofectin treatment and in analogy, none of the inactive ODN induced proliferation after lipofectin treatment. Among PBMC, ODN-responding cells were identified as predominantly Bu-1, immunoglobulin and major histocompatibility complex class II expressing cells, while CD3 expressing cells were not responding. Using magnetic cell separation of Bu-1 expressing cells prior to culture it was found that Bu-1 depleted cells did not proliferate upon ODN stimulation while the Bu-1 enriched cells were able to proliferate upon this stimulus. Taken together, among ODN in the present panel, only phosphorothioate ODN induced proliferation of PBMC. Responses were induced regardless of the presence of CpG-motifs and were not influenced by addition of lipofectin. Amid the chicken PBMC, predominantly cells of a B-cell phenotype proliferated in response to ODN stimulation and they were able to respond to this stimulus without the presence of other cell types.

Immune-related gene expression in response to H11N9 low pathogenic avian influenza virus infection in chicken and Pekin duck peripheral blood mononuclear cells

The duck and chicken are important hosts of avian influenza virus (AIV) with distinctive responses to infection. Frequently, AIV infections in ducks are asymptomatic and long-lasting in contrast to the clinically apparent and transient infections observed in chickens. These differences may be due in part to the host response to AIV infection. Using real-time quantitative PCR, we examined the expression of immune-related genes in response to low pathogenic AIV H11N9 infection in peripheral blood mononuclear cells (PBMC) isolated from the blood of chickens and Pekin ducks. While chicken PBMC expressed IL-1β and IL-6 at high levels similar to mammalian species, duck PBMC expression levels were minimal or unchanged. Similarly, duck IFN-β expression was nearly unaffected, whereas chicken expression was highly upregulated. Chicken IFN-γ was expressed to higher levels than duck IFN-γ, while IFN-α was expressed similarly by both species. IL-2 was elevated early in infection in duck PBMC, but returned to baseline levels by the end of the experiment; in contrast, IL-2 was weakly induced in chicken PBMC at late time points. TLR-7 and MHC class I molecule expressions were conserved between species, whereas duck MHC class II expression was downregulated and chicken expression was unchanged. These results show distinct PBMC expression patterns of pro-inflammatory cytokines and IFNs between species. The differences in pro-inflammatory cytokine and IFN expression reflect the asymptomatic and lasting infection observed in ducks and the tendency towards clinical signs and rapid clearance seen in chickens. These results highlight important differences in the host response to AIV of two species thought to be critical in the genesis and maintenance of epidemic strains of AIV.

The selective Dectin-1 agonist, curdlan, induces an oxidative burst response in chicken heterophils and peripheral blood mononuclear cells

A critical component of host innate immunity is recognition of pathogen-associated molecular patterns (PAMPs) by pattern recognition receptors (PRRs). Dectin-1 is the primary PRR for exogenous β-glucan, a component of fungal and bacterial cell walls. A previous study conducted in our laboratory demonstrated that administration of β-glucan as a feed additive resulted in increased innate immune function of neonatal chickens, suggesting that chickens possess a Dectin-1-like β-glucan receptor. In the present study, we demonstrated that heterophils and peripheral blood mononuclear cells (PBMCs) from day-old chicks had a significant increase in the generation of reactive oxygen species (ROS) following stimulation with the Dectin-1 specific agonist, curdlan. Pretreatment of heterophils and PBMCs with laminarin, a β-glucan receptor blocking agent and specific inhibitor of Dectin-1 activity, significantly reduced the curdlan-induced ROS production. Together these data provide evidence for the first time of the presence of a functional Dectin-1-like β-glucan receptor in chicken heterophils and PBMCs.

Baculovirus up-regulates antiviral systems and induces protection against infectious bronchitis virus challenge in neonatal chicken

In this study, the Antheraea pernyi nuclear polyhedrosis virus (ApNPV), a member of the baculovirus family, is evaluated for its stimulation of chicken peripheral blood mononuclear cells (PBMC) and macrophage cell line HD 11 in vitro, and protection against infectious bronchitis virus (IBV) in neonatal chickens in vivo. This study showed that ApNPV significantly enhanced inflammatory cytokine mRNA expression in chicken PBMC and HD 11 cells through the clathrin-dependent endocytic and endosomal maturation pathway, and up-regulated nitric oxide production in HD 11 cells. Furthermore, it was identified that budded virus (BV) can induce antiviral effects in HD 11 cells contrary to occlusion-derived virus (ODV). These results indicate that immunostimulatory BV of ApNPV can stimulate the innate immune activities and enhance the resistance to infectious virus of neonatal chickens.

Cytokine Expression in Chicken Peripheral Blood Mononuclear Cells after In Vitro Exposure to Salmonella enterica serovar Enteritidis

Cytokines are secreted proteins involved with cell recruitment and regulation of both innate and adaptive immune responses. They are essential for an effective host immune response to pathogens. The objective of this study was to determine the effect of Salmonella enterica serovar Enteritidis (S. Enteritidis) exposure and genetic line on cytokine mRNA expression level of cultured chicken peripheral blood mononuclear cells (PBMC). Interleukin-2, interleukin-6 (IL-6), CXCLi2, and transforming growth factor-β4 (TGF-B4) messenger ribonucleic acid expression was measured by quantitative reverse transcription-PCR assays in PBMC from 3 chicken lines (broiler, Leghorn, Fayoumi) after in vitro exposure to S. Enteritidis. The PBMC were isolated from uninfected birds and cultured overnight. The next day, live pathogenic S. Enteritidis was added to half of the cultures. All cultures were harvested after 2 or 4 h of exposure. Exposure to S. Enteritidis downregulated IL-6, CXCLi2, and TGF-β4 but not interleukin-2 mRNA expression. No significant genetic line or exposure time effects were detected. These findings demonstrate that exposure of chicken PBMC to S. Enteritidis can induce a rapid change in both proinflammatory (IL-6, CXCLi2) and antiinflammatory (TGF-β4) cytokine gene expression.