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Quintana AM Landolt GA Annis KM Hussey GS 《Veterinary immunology and immunopathology》2011,140(3-4):226-236
Our understanding of innate immunity within the equine respiratory tract is limited despite growing evidence for its key role in both the immediate defense and the shaping of downstream adaptive immune responses to respiratory disease. As the first interface to undergo pathogen invasion, the respiratory epithelium is a key player in these early events and our goal was to examine the innate immune characteristics of equine respiratory epithelia and compare them to an in vitro equine respiratory epithelial cell model cultured at the air-fluid interface (AFI). Respiratory epithelial tissues, isolated epithelial cells, and four-week old cultured differentiated airway epithelial cells collected from five locations of the equine respiratory tract were examined for the expression of toll-like receptors (TLRs) and host defense peptides (HDPs) using conventional polymerase chain reaction (PCR). Cultured, differentiated, respiratory epithelial cells and freshly isolated respiratory epithelial cells were also examined for the expression of TLR3, TLR9 and major histocompatibility complex (MHC) class I and class II using fluorescence-activated cell sorting (FACS) analysis. In addition, cytokine and chemokine profiles from respiratory epithelial tissues, freshly isolated respiratory epithelial cells, and cultured, differentiated, epithelial cells from the upper respiratory tract were examined using real-time PCR. We found that respiratory epithelial tissues and isolated epithelial cells expressed TLRs 1-4 and 6-10 as well as HDPs, MxA, 2'5' OAS, β-defensin-1, and lactoferrin. In contrast, epithelial cells cultured at the AFI expressed TLRs 1-4 and 6 and 7 as well as MxA, 2'5' OAS, β-defensin-1, but had lost expression of TLRs 8-10 and lactoferrin. In addition, MHC-I and MHC-II surface expression decreased in epithelial cells cultured at the AFI compared to isolated epithelial cells whereas TLR3 and TLR9 were expressed at similar levels. Lastly, we found that equine respiratory epithelial cells express an array of pro-inflammatory, antiviral and regulatory cytokines and that after four weeks of in vitro growth conditions, equine respiratory epithelial cells cultured at the AFI retained expression of GM-CSF, IL-10, IL-8, TGF-β, TNF-α, and IL-6. In summary, we describe the development of an in vitro equine respiratory epithelial cell culture model that is morphologically similar to the equine airway epithelium and retains several key immunological properties. In the future this model will be a used to study equine respiratory viral pathogenesis and cell-to-cell interactions. 相似文献
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Ignacio G Nordone S Howard KE Dean GA 《Veterinary immunology and immunopathology》2005,106(3-4):229-237
Toll-like receptors (TLRs) are germline-encoded pattern recognition receptors (PRRs) that activate the innate immune system. While it is clear that TLRs are important in the immune response against pathogens, they may also be exploited by some pathogens. Our objective is to determine whether feline immunodeficiency virus (FIV) infection affects TLR expression or function thereby resulting in innate immune dysfunction. To this end, we cloned partial sequences for feline TLRs 1--3, 5--8, and developed real-time PCR assays to quantify feline TLRs 1--9. TLR expression was quantified in normal cat lymphoid tissues, purified lymphocyte subsets, and FIV-infected cell lines. Different expression patterns of TLRs were found in spleen, mesenteric lymph node, retropharyngeal lymph node, thymus, intestinal intraepithelial lymphocytes, and lamina propria lymphocytes. B lymphocytes, CD4+ T cells, and CD8+ T cells all expressed TLRs 2--5, 7--9; however, the relative levels of expression varied among lymphocyte phenotypes. Infection of cell lines with FIV resulted in altered TLR expression levels that differed depending on cell type. These results demonstrate that tissue distribution of TLRs is associated with the immunological role of a particular tissue, that lymphocytes may also express these 'innate immune' receptors, and that FIV infection can alter TLR expression. 相似文献
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Kwong LS Parsons R Patterson R Coffey TJ Thonur L Chang JS Russell G Haig D Werling D Hope JC 《Veterinary immunology and immunopathology》2011,139(2-4):313-318
Host recognition of conserved pathogen-associated molecular patterns (PAMPs) and their interactions with pattern-recognition receptors, including the Toll-like receptors (TLR) is essential for innate immune response induction. The TLR1 family (TLR1, 2, 6 and 10) is involved in the recognition of gram-positive and gram-negative bacteria and heterodimers of TLR1 or TLR6 with TLR2 are crucial for the identification of several PAMPs. Studies on cell surface expression of TLR in ruminants are hampered by the lack of specific antibodies and no convincingly cross-reactive anti-human antibodies have been described so far. We describe herein four antibodies which recognise bovine TLR2. Differences in TLR2 expression were evident on bovine antigen presenting cells with high level expression on peripheral blood monocytes and monocyte-derived macrophages. Lower levels of expression were evident on dendritic cell populations derived in vitro and ex vivo, and on alveolar macrophages. One of the antibodies recognised TLR2 expression on ovine peripheral blood monocytes. The identification of antibodies specific for bovine and ovine TLR2 will facilitate studies of the role of this important PRR in the initiation of immune responses to important pathogens. 相似文献
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《Veterinary immunology and immunopathology》2015,163(3-4):157-163
Cells of the adaptive immune system express Toll-like receptors (TLRs) and are able to respond to TLR ligands. With this in mind, the goal of the current study was to determine the expression of antiviral response genes in the cells of the chicken bursa of Fabricius (BF) to stimulation with TLR ligands. We investigated initially the response of bursal B cells to CpG-ODN, lipopolysaccharide (LPS) and poly(I:C) treatment. The expression level of type I interferons (IFNs) and interferon regulatory factor 7 (IRF7) did not differ between CpG-ODN and LPS treated groups compared to the non-stimulated cells. Poly(I:C) was the only TLR ligand, which has induced significant expression of antiviral innate immune response genes from bursal cells. Further in vitro and in vivo studies need to examine the efficacy of these antiviral responses against avian viruses. 相似文献
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Xuming Hu Wencai Xu Aijian Qin Genghua Wu Kun Qian Hongxia Shao Jianqiang Ye 《Veterinary research communications》2014,38(2):149-156
Marek’s disease virus (MDV) is a highly oncogenic alpha-herpesvirus that causes T cell immune suppression and malignant lymphomas in chickens. Toll-like receptor (TLR) plays a dominant role in antiviral T cell immunity. However, it is unclear whether MDV induced T cell immunity is associated with TLR-mediated immunity. In this study, the expression of 28 host genes that are involved in TLR-mediated immunity and MHC-medicated T cell immunity was evaluated in chicken thymus at 7, 14, 21 and 28 days post-infection (dpi). Our results demonstrated that 24 host immune-related genes were upregulated during MDV infection at 7 dpi; however, the expression of most of these genes decreased at 21 and 28 dpi. Notably, a positive correlation was found between the down-regulation of CD4, CD8 and TLR3 signals but not the MyD88-dependent TLR pathway. The present study expanded our knowledge of host immune responses against MDV infection and our results might provide a clue that MDV may interfere with T cell immune response through TLR3 signals. 相似文献
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Jinfeng Liang Jia FuHaihong Kang Jian LinQinghua Yu Qian Yang 《Veterinary immunology and immunopathology》2013
Dendritic cells (DCs) are crucial for initiation of both innate and adaptive immune responses. TLR ligands combine with Toll-like receptors (TLRs) expressed on the DC surface and induce DC maturation. The potential effect of three types of TLR ligands (Bacillus subtilis (B. subtilis) spores, polyinosinic–polycytidylic acid and CpG oligodeoxynucleotides) on chicken bone marrow-derived DCs (chBM-DCs) maturation was studied. The chBM-DCs cultured in presence of recombinant chicken granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4 displayed the typical morphology of DCs after 7 days of culture. These immature chBM-DCs up-regulated the expression of MHC-II and of the putative CD11c, but had yet low to moderate levels of the CD40 and CD86 co-stimulatory molecules. After stimulation by the TLR ligands, the chBM-DCs displayed a more mature morphologic phenotype, significantly increased the CD40 and CD86 cell surface expression levels and gained the ability to stimulate proliferation of naive T cells in the allogeneic mixed lymphocyte reaction, compared to the immature chBM-DCs. In conclusion, our data demonstrated that all three TLR ligands were strong stimuli for driving chBM-DCs maturation in vitro, with B. subtilis spores being the most efficient. 相似文献
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To evaluate effects of treatment with pathogen-associated molecular patterns (PAMPs) on toll-like receptor (TLR), MHC II, B7 and cytokine expression, pig monocytes and monocyte-derived DCs (moDCs) were treated with LPS, CpG, lipoteichoic acid (LTA), poly IC or peptidoglycan (Pep). Monocytes and moDCs treated with LPS, CpG, LTA, poly IC or Pep altered expression of at least one TLR (4, 5 and 9) and up-regulated MHC II and/or B7. The mRNA for IL-4 was not detected after any treatment. Treatment with LPS or LTA tended to up-regulate mRNA for TLR 4, Th-1 (IFN-gamma and IL-12p35) and Th-2 cytokines (IL-10 and IL-13). Poly IC or CpG tended to up-regulate TLR 9 and Th-1 cytokines. Porcine monocytes and moDCs like those of humans and mice responded to microbial PAMPs by altering TLR expression, up-regulating MHC II and B7 and altering cytokine expression toward Th-1 and/or Th-2, which may steer immune response. Hence, porcine moDCs and monocytes are likely able to discriminate between microorganisms using TLRs which determine cytokine expression and immune response bias. 相似文献
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M.S. Marin S. Quintana C. Faverín M.R. Leunda A.C. Odeón S.E. Pérez 《Research in veterinary science》2014
The involvement of Toll-like receptors (TLRs) in bovine herpesvirus types 1 (BoHV-1) and 5 (BoHV-5) infections has not been analyzed. In this study, the role of TLR signaling on virus replication was investigated. Blood leukocytes consistently express TLRs. Thus, our approach was to study in vitro the effects of agonist stimulation of TLRs expressed by peripheral blood leukocytes on BoHV-1 and BoHV-5 replication. Furthermore, the patterns of TLRs 3, 7–9 expression on virus-infected-bovine leukocytes were analyzed. Only Imiquimod (TLR7/8 agonist) showed anti-viral activity on infected MDBK cells. This is the first evidence that the timely activation of TLR7/8 signaling is effective in impairing BoHV-1 and 5 replication, thereby providing an experimental indication that Imiquimod may be a promising immune modulator. This work describes, for the first time, the expression patterns of TLRs in BoHV-1- or BoHV-5-infected-bovine leukocytes, suggesting the involvement of TLR7 and TLR9 in the recognition of these viruses. 相似文献
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Toll-like receptors (TLRs), a family of transmembrane and cytosolic proteins, detect microbial patterns, initiating innate immune responses in various organisms. Although they are abundant, genetic characterization and functional differences of TLRs in economically important avian species such as chickens and turkeys have not been investigated in detail. In this study, the putative TLR5 coding region from turkey genome was sequenced, and its homology to other vertebrate species was analyzed. Secondary structure analysis revealed protein motifs typical of the chicken TLR5 protein structure, with 97% amino acid identity between them. mRNA expression profiling in adult turkeys revealed abundant TLR5 expression in a broad range of tissues. Stimulation with the TLR5 ligand flagellin resulted in the production of the inflammatory mediators interleukin (IL)-1beta, IL-6, and nitric oxide in peripheral blood mononuclear cells. To our knowledge, this is the first complete turkey TLR5 coding DNA sequence reported in sequence databases. 相似文献
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To investigate the effects of different doses of compound Chinese herbal medicinal polysaccharides (cCHMPS) on TLR4 and downstream MyD88 dependent signal transduction pathway components in chicken lymphocytes of different MHC B-LβⅡ genotypes,PCR-SSCP technique was applied to group layer according to different MHC B-LβⅡ genotypes.The peripheral blood lymphocytes of chicken with different MHC B-LβⅡ genotypes were collected,and added with 100,75,50 and 0 μg/mL cCHMPS (high,middle and low dose groups and control group),respectively,then co-culturing for 16,24,32 and 48 h.The expression of TLR4,MYD88 and TRAF-6 mRNA were detected using Real-time PCR method.The results showed that compared with control group,cCHMPS could significantly improve the expression levels of TLR4,MYD88 and TRAF-6 mRNA of different MHC B-Lβ Ⅱ genotypes chickens (P < 0.05);The expression levels of TLR4,MYD88 and TRAF-6 mRNA of AA genotype chicken lymphocyte in middle and low dose groups were higher than those of high dose group (except TLR4 gene cultured for 16 h);The expression of TLR4,MyD88 and TNAF-6 mRNA of BB genotype in high dose group were higher than those of other dose groups (except TLR4 gene cultured for 32 and 48 h);The expression of TLR4,MyD88 and TNAF-6 mRNA of BC genotype in low dose were higher than that of other dose groups (except TLR4 gene cultured for 16 h).There results indicated that cCHMPS played an important role in the body’s immune regulatory mechanism by binding to TLR4 in the surface of lymphocytes,activating the downstream MyD88-dependent signal transduction pathway,regulating cellular immunity,and cCHMPS optimum immunomodulatory does were different in each MHC B-Lβ Ⅱ genotype chickens. 相似文献
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Hosoya S Villena J Shimazu T Tohno M Fujie H Chiba E Shimosato T Aso H Suda Y Kawai Y Saito T Alvarez S Ikegami S Itoh H Kitazawa H 《Veterinary research》2011,42(1):111
ABSTRACT: This study analyzed the functional expression of TLR3 in various gastrointestinal tissues from adult swine and shows that TLR3 is expressed preferentially in intestinal epithelial cells (IEC), CD172a+CD11R1high and CD4+ cells from ileal Peyer's patches. We characterized the inflammatory immune response triggered by TLR3 activation in a clonal porcine intestinal epitheliocyte cell line (PIE cells) and in PIE-immune cell co-cultures, and demonstrated that these systems are valuable tools to study in vitro the immune response triggered by TLR3 on IEC and the interaction between IEC and immune cells. In addition, we selected an immunobiotic lactic acid bacteria strain, Lactobacillus casei MEP221106, able to beneficially regulate the anti-viral immune response triggered by poly(I:C) stimulation in PIE cells. Moreover, we deepened our understanding of the possible mechanisms of immunobiotic action by demonstrating that L. casei MEP221106 modulates the interaction between IEC and immune cells during the generation of a TLR3-mediated immune response. 相似文献
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Porcherie A Cunha P Trotereau A Roussel P Gilbert FB Rainard P Germon P 《Veterinary research》2012,43(1):14
ABSTRACT: Escherichia coli is a frequent cause of clinical mastitis in dairy cows. It has been shown that a prompt response of the mammary gland after E. coli entry into the lumen of the gland is required to control the infection, which means that the early detection of bacteria is of prime importance. Yet, apart from lipopolysaccharide (LPS), little is known of the bacterial components which are detected by the mammary innate immune system. We investigated the repertoire of potential bacterial agonists sensed by the udder and bovine mammary epithelial cells (bMEC) during E. coli mastitis by using purified or synthetic molecular surrogates of bacterial agonists of identified pattern-recognition receptors (PRRs). The production of CXCL8 and the influx of leucocytes in milk were the readouts of reactivity of stimulated cultured bMEC and challenged udders, respectively. Quantitative PCR revealed that bMEC in culture expressed the nucleotide oligomerization domain receptors NOD1 and NOD2, along with the Toll-like receptors TLR1, TLR2, TLR4, and TLR6, but hardly TLR5. In line with expression data, bMEC proved to react to the cognate agonists C12-iE-DAP (NOD1), Pam3CSK4 (TLR1/2), Pam2CSK4 (TLR2/6), pure LPS (TLR4), but not to flagellin (TLR5). As the udder reactivity to NOD1 and TLR5 agonists has never been reported, we tested whether the mammary gland reacted to intramammary infusion of C12-iE-DAP or flagellin. The udder reacted to C12-iE-DAP, but not to flagellin, in line with the reactivity of bMEC. These results extend our knowledge of the reactivity of the bovine mammary gland to bacterial agonists of the innate immune system, and suggest that E. coli can be recognized by several PRRs including NOD1, but unexpectedly not by TLR5. The way the mammary gland senses E. coli is likely to shape the innate immune response and finally the outcome of E. coli mastitis. 相似文献
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The objective of this study was to identify the effect of in vitro lipopolysaccharide (LPS) treatment on regulatory T cells (Tregs) from chickens. Tregs had approximately 30-fold higher TLR 2-type 2 and six-fold higher TLR 4 mRNA content than CD4+CD25- cells. Tregs were treated with either 0 or 1 μg/ml LPS for 0, 2, and 4d. LPS treatment increased the IL-2 mRNA amount in Tregs at 2 and 4d post-LPS treatment. LPS treatment increased the IL-10 mRNA amount in Tregs at 4d post-LPS treatment. The total live cell numbers were approximately two-fold higher at 2d and three-fold higher at 4d in the 1 μg/ml LPS-treated groups than in the 0 μg LPS-treated controls. LPS treatment abrogated suppressive properties of Tregs at 2d post-LPS treatment. At 4d post-LPS treatment, Tregs became supersuppressive. In conclusion, chicken Tregs are differentially activated to facilitate immune response during the early stage of inflammation and to facilitate immune suppression at a later stage of inflammation. 相似文献
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Subharat S Shu D de Lisle GW Buddle BM Wedlock DN 《Veterinary immunology and immunopathology》2012,145(1-2):471-478
Johne's disease caused by Mycobacterium avium subsp. paratuberculosis (MAP), is a chronic enteric disease of cattle. The mechanism how MAP can co-exist in the gastro-intestinal tract despite a massive infiltration of immune cells is not known. Toll-like receptors (TLRs) are known to play an important role in both innate and acquired immune responses but it is unclear what role different TLRs play in response to MAP. In this study, 38 cull cows from herds infected with MAP were classified into four groups, based on MAP culture from gut tissues and histopathological lesion scores. The expression of TLR1, 2 and 4 mRNA from MAP antigen-stimulated mesenteric lymph node (MLN) cultures and peripheral blood mononuclear cells (PBMCs) and in the MLN and ileum tissues of these animals was determined. MAP antigen-specific expression of TLR1 in MLN and PBMC was significantly lower in the MAP-infected groups than the non-infected control group, suggesting that in MAP-infected animals there is impairment in the up-regulation of TLR1 in response to MAP antigen. TLR4 expression in MLN tissues was significantly higher in the severely infected group than the control group suggesting up-regulation of endogenous TLR4 expression at a site of MAP infection in animals severely affected with Johne's disease. A preliminary screening of TLR1, 2 and 4 in the cull cows revealed the presence of polymorphisms in TLR1 and TLR2. In summary, one mechanism how MAP may subvert the immune system is that there is an apparent lack of recognition of MAP antigens as foreign by TLR1 in MAP-infected cows. 相似文献