Experimental model of equine alveolar macrophage stimulation with TLR ligands

Waldschmidt, Ingrid; Pirottin, Dimitri; Art, Tatiana; Audigié, Fabrice; Bureau, Fabrice; Tosi, Irène; Abbas, Sophie El; Farnir, Frédéric; Richard, Éric; Dupuis, Mari-Capucine

doi:10.1016/j.vetimm.2013.05.017

Cited by 8 publications

(6 citation statements)

References 34 publications

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Section: Discussionmentioning

confidence: 99%

“…Although the LPS response of equine AMs has been subject to previous study by others, largely justified by the hypothesised role of inhaled endotoxin in certain equine environmental airway diseases , the data presented here are the most comprehensive to date. LPS‐induced upregulation of many of the genes identified has previously been reported in a variety of species, including the horse . In comparison with equine monocytes, in which LPS seems to activate mainly targets of the MyD88 pathway , our data revealed LPS‐induced activation of both MyD88 ( TNF , IL1 , IL6 , IL10 ) and TRIF ( IFNB , CCL5 ) pathways in equine AMs, a finding which may reflect a degree of adaptation to the unique airway microenvironment.…”

Section: Discussionmentioning

confidence: 99%

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Comparative transcriptome analysis of equine alveolar macrophages

Karagianni

Kapétanovic

Summers

et al. 2016

Equine Veterinary Journal

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SummaryReasons for performing studyAlveolar macrophages (AMs) are the first line of defence against pathogens in the lungs of all mammalian species and thus may constitute appropriate therapeutic target cells in the treatment and prevention of opportunistic airway infections. Therefore, acquiring a better understanding of equine macrophage biology is of paramount importance in addressing this issue in relation to the horse.ObjectivesTo compare the transcriptome of equine AMs with that of equine peritoneal macrophages (PMs) and to investigate the effect of lipopolysaccharide (LPS) on equine AM.Study designGene expression study of equine AMs.MethodsCells from both bronchoalveolar and peritoneal lavage fluid were isolated from systemically healthy horses that had been submitted to euthanasia. Cells were cryopreserved. RNA was extracted and comparative microarray analyses were performed in AMs and PMs, and in AMs treated and untreated with LPS. Comparisons with published data derived from human AM studies were made, with particular focus on LPS‐induced inflammatory status.ResultsThe comparison between AMs and PMs revealed the differential basal expression of 451 genes. Gene expression analysis revealed an alternative (M2) macrophage polarisation profile in AMs and a hybrid macrophage activation profile in PMs, a phenomenon potentially attributable to a degree of induced endotoxin tolerance. The gene expression profile of equine AMs following LPS stimulation revealed significant changes in the expression of 240 genes, including well‐known upregulated inflammatory genes. This LPS‐induced gene expression profile of equine AMs more closely resembles that of human rather than murine macrophages.ConclusionsThis study improves current understanding of equine macrophage biology. These data suggest that the horse may represent a suitable animal model for the study of human macrophage‐associated lung inflammation and data derived from human macrophage studies may have significant relevance to the horse.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Comparative transcriptome analysis of equine alveolar macrophages

Karagianni

Kapétanovic

Summers

et al. 2016

Equine Veterinary Journal

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“…Microbiopsies of the gluteus medius muscle were obtained as previously described [ 28 ]. This tissue served as calibrator/control tissue for the TLR expression as previously described [ 28 ] since muscle tissue is known to have the lowest TLR expression within the body tissues [ 29 ].…”

Section: Methodsmentioning

confidence: 99%

“…Only RNA samples with an RNA integrity number (RIN) ≥ 9 and a 28S:18S ratio of > 2:1 qualified as “good quality” RNA and were used for real-time PCR. cDNA were generated as previously described [ 28 ]. Gene expression of TLR1-9, fibroblast growth factor-1 (FGF-1), fibroblast specific protein-1 (FSP-1), E-cadherin, keratin19, IFN-β, TNF-α, IL-6 and CXCL8 was quantified by real-time PCR using Absolute Blue qPCR SYBR Green Mix (Fisher Scientific, Aalst, Belgium) on an Applied Biosystems 7900 HT thermocycler (Applied Biosystems, Carlsbad, CA, USA).…”

Section: Methodsmentioning

confidence: 99%

The innate immune response of equine bronchial epithelial cells is altered by training

Frellstedt

Gosset

Kervoaze

et al. 2015

Vet Res

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Respiratory diseases, including inflammatory airway disease (IAD), viral and bacterial infections, are common problems in exercising horses. The airway epithelium constitutes a major physical barrier against airborne infections and plays an essential role in the lung innate immune response mainly through toll-like receptor (TLR) activation. The aim of this study was to develop a model for the culture of equine bronchial epithelial cells (EBEC) in vitro and to explore EBEC innate immune responses in trained horses. Bronchial epithelial biopsies were taken from 6 adult horses during lower airway endoscopy. EBEC were grown in vitro by an explant method. The innate immune response of EBEC was evaluated in vitro by treatment with TLR ligands. TLR3 is the most strongly expressed TLR at the mRNA level in EBEC and stimulation of EBEC with Poly(I:C), an analog of viral dsRNA, triggers a strong secretion of IFN-β, TNF-α, IL-6 and CXCL8. We further evaluated the EBEC innate immune response in horses that underwent a 4-month-training program. While training had no effect on TLR mRNA expression in EBEC as well as in bronchial biopsies, it increased the production of IFN-β after stimulation with a TLR3 ligand and decreased the secretion of TNF-α and IL-6 after stimulation with a TLR2 and TLR3 ligand. These findings may be implicated in the increased risk for viral and bacterial infections observed in sport horses. Altogether, we report a successful model for the culture of EBEC that can be applied to the investigation of pathophysiologic conditions in longitudinal studies.

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“…Expression of TLR2 and TLR6 was demonstrated in porcine alveolar macrophages by Western blot and using antibodies against these receptors, their relevance in the sensing of Mycoplasma hyopneumoniae was shown [17]. Also in the horse, TLR2 expression was detected by RT-PCR in alveolar macrophages [31], as well as in respiratory epithelia [32] and PBMCs [33]. …”

Section: Tlr2mentioning

confidence: 99%

Targeting TLR2 for Vaccine Development

Basto

Leitão

2014

Journal of Immunology Research

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Novel and more effective immunization strategies against many animal diseases may profit from the current knowledge on the modulation of specific immunity through stimulation of innate immune receptors. Toll-like receptor (TLR)2-targeting formulations, such as synthetic lipopeptides and antigens expressed in fusion with lipoproteins, have been shown to have built-in adjuvant properties and to be effective at inducing cellular and humoral immune mechanisms in different animal species. However, contradictory data has arisen concerning the profile of the immune response elicited. The benefits of targeting TLR2 for vaccine development are thus still debatable and more studies are needed to rationally explore its characteristics. Here, we resume the main features of TLR2 and TLR2-induced immune responses, focusing on what has been reported for veterinary animals.

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Experimental model of equine alveolar macrophage stimulation with TLR ligands

Cited by 8 publications

References 34 publications

Comparative transcriptome analysis of equine alveolar macrophages

Comparative transcriptome analysis of equine alveolar macrophages

The innate immune response of equine bronchial epithelial cells is altered by training

Targeting TLR2 for Vaccine Development

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