Effect of hay dust extract and cyathostomin antigen stimulation on cytokine expression by PBMC in horses with recurrent airway obstruction

Lanz, Simone; Gerber, Vinzenz; Marti, Eliane; Rettmer, Helen; Klukowska‐Rötzler, Jolanta; Gottstein, Bruno; Matthews, J. B.; Pirie, Scott; Hamza, Eman

doi:10.1016/j.vetimm.2013.07.005

Cited by 22 publications

(40 citation statements)

References 55 publications

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“…This study was approved by the Animal Experimentation Committee of the Canton of Bern, Switzerland (BE33/07, BE58/10 and BE10/13). The sample collection was previously described in detail in an earlier publication [49]. …”

Section: Methodsmentioning

confidence: 99%

“…This study is based on the results generated in a previously reported experiment and therefore all methods in detail are described elsewhere [49, 50, 56]. Briefly, blood samples were collected from 41 adult Warmblood horses (free of RAO) from three cohorts: two half-sibling families (Fam1: 7 horses, Fam2: 9 horses) and a group of unrelated horses (Un: 25 horses).…”

Section: Methodsmentioning

confidence: 99%

“…They include cells that are involved in both the innate (monocytes, dendritic cells) and the acquired (lymphocytes) immune response systems. Therefore, PBMCs to a certain extent reflect the immune system status [48] and are widely used in systemic immune response studies [49–55]. …”

Section: Introductionmentioning

confidence: 99%

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LPS-induced modules of co-expressed genes in equine peripheral blood mononuclear cells

et al. 2017

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BackgroundLipopolysaccharide (endotoxin, LPS) is a strong inducer of the innate immune response. It is widespread in our environment, e.g. in house dust and contributes to asthma. Compared to humans, horses are even more sensitive to LPS. However, data on LPS effects on the equine transcriptome are very limited. Using RNA-seq we analysed LPS-induced differences in the gene expression in equine peripheral blood mononuclear cells at the gene and gene-network level in two half-sib families and one group of unrelated horses.Results24 h-LPS challenge of equine immune cells resulted in substantial changes in the transcriptomic profile (1,265 differentially expressed genes) showing partial overlap with human data. One of the half-sib families showed a specific response different from the other two groups of horses. We also identified co-expressed gene modules that clearly differentiated 24 h-LPS- from non-stimulated samples. These modules consisted of 934 highly interconnected genes and included genes involved in the immune response (e.g. IL6, CCL22, CXCL6, CXCL2), however, none of the top ten hub genes of the modules have been annotated as responsive to LPS in gene ontology.ConclusionsUsing weighted gene co-expression network analysis we identified ten co-expressed gene modules significantly regulated by in vitro stimulation with LPS. Apart from 47 genes (5%) all other genes highly interconnected within the most up- and down-regulated modules were also significantly differentially expressed (FDR < 0.05). The LPS-regulated module hub genes have not yet been described as having a role in the immune response to LPS (e.g. VAT1 and TTC25).Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-3390-y) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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LPS-induced modules of co-expressed genes in equine peripheral blood mononuclear cells

et al. 2017

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“…Briefly, blood was collected from jugular vein into sodium-heparin-containing vacutainers [54]. Within 1–6 h serum was separated from cells via centrifugation 1800 g for 20 min.…”

Section: Methodsmentioning

confidence: 99%

Novel equine tissue miRNAs and breed-related miRNA expressed in serum

et al. 2016

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BackgroundMiRNAs regulate multiple genes at the post-transcriptional level and therefore play an important role in many biological processes. It has been suggested that miRNA exported outside the cells contribute to inter-cellular communication. Consequently, circulating miRNAs are of particular interest and are promising biomarkers for many diseases. The number of miRNAs annotated in the horse genome is much lower compared to model organisms like human and mouse. We therefore aimed to identify novel equine miRNAs for tissue types and breed in serum.ResultsWe analysed 71 small RNA-seq libraries derived from nine tissues (gluteus medius, platysma, masseter muscle, heart, liver, cartilage, bone, total blood and serum) using miRDeep2 and miRdentify tools. Known miRNAs represented between 2.3 and 62.9 % of the reads in 71 libraries. A total of 683 novel miRNAs were identified. Breed and tissue type affected the number of miRNAs detected and interestingly, affected its average intensity. A total of 50 miRNAs in serum proved to be potential biomarkers to differentiate specific breed types, of which miR-122, miR-200, miR-483 were over-expressed and miR-328 was under-expressed in ponies compared to Warmbloods. The different miRNAs profiles, as well as the differences in their expression levels provide a foundation for more hypotheses based on the novel miRNAs discovered.ConclusionsWe identified 683 novel equine miRNAs expressed in seven solid tissues, blood and serum. Additionally, our approach evidenced that such data supported identification of specific miRNAs as markers of functions related to breeds or disease tissues.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-3168-2) contains supplementary material, which is available to authorized users.

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“…Duplicate cDNA samples were amplified for 45 cycles (15 s at 95 • C and 60 s at 60 • C) in an AB7300 real-time PCR instrument (Applied Biosystems, CA, USA) using Maxima Probe qPCR Mastermix (Fermentas). Cycle threshold (Ct) values of cytokine gene expression were normalized to the 18s rRNA content ( CT) as described in Lanz et al (2013). No significant difference was detected in any of the measured parameters in the un-stimulated cells between the groups of horses.…”

Section: Stimulation Of Pbmcmentioning

confidence: 99%

Developing a preventive immunization approach against insect bite hypersensitivity using recombinant allergens: A pilot study

Jónsdóttir

Hamza

Janda

et al. 2015

Veterinary Immunology and Immunopathology

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Effect of hay dust extract and cyathostomin antigen stimulation on cytokine expression by PBMC in horses with recurrent airway obstruction

Cited by 22 publications

References 55 publications

LPS-induced modules of co-expressed genes in equine peripheral blood mononuclear cells

LPS-induced modules of co-expressed genes in equine peripheral blood mononuclear cells

Novel equine tissue miRNAs and breed-related miRNA expressed in serum

Developing a preventive immunization approach against insect bite hypersensitivity using recombinant allergens: A pilot study

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