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BMC veterinary research2016; 12(1); 117; doi: 10.1186/s12917-016-0742-4

Release kinetics of tumor necrosis factor-α and interleukin-1 receptor antagonist in the equine whole blood.

Abstract: Horses are much predisposed and susceptible to excessive and acute inflammatory responses that cause the recruitment and stimulation of polymorphnuclear granulocytes (PMN) together with peripheral blood mononuclear cells (PBMC) and the release of cytokines. The aim of the study is to develop easy, quick, cheap and reproducible methods for measuring tumor necrosis factor alpha (TNF-α) and interleukin-1 receptor antagonist (IL-1Ra) in the equine whole blood cultures ex-vivo time- and concentration-dependently. Results: Horse whole blood diluted to 10, 20 and 50 % was stimulated with lipopolysaccharide (LPS), PCPwL (a combination of phytohemagglutinin E, concanavalin A and pokeweed mitogen) or equine recombinant TNF-α (erTNF-α). TNF-α and IL-1Ra were analyzed in culture supernatants, which were collected at different time points using specific enzyme-linked immunosorbent assays (ELISA). Both cytokines could be detected optimal in stimulated 20 % whole blood cultures. TNF-α and IL-1Ra releases were time-dependent but the kinetic was different between them. PCPwL-induced TNF-α and IL-1Ra release was enhanced continuously over 24-48 h, respectively. Similarly, LPS-stimulated TNF-α was at maximum at time points between 8-12 h and started to decrease thereafter, whereas IL-1Ra peaked later between 12-24 h and rather continued to accumulate over 48 h. The equine recombinant TNF-α could induce also the IL-1Ra release. Conclusions: Our results demonstrate that similar to PCPwL, LPS stimulated TNF-α and IL-1Ra production time-dependently in whole blood cultures, suggesting the suitability of whole blood cultures to assess the release of a variety of cytokines in health and diseases of horse.
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Publication Date: 2016-06-17 PubMed ID: 27316332PubMed Central: PMC4912716DOI: 10.1186/s12917-016-0742-4Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research aims to devise a cost-effective and reliable method to measure certain inflammatory compounds, specifically TNF-α (Tumor Necrosis Factor Alpha) and IL-1Ra (Interleukin-1 Receptor Antagonist), in the blood of horses. These compounds play a significant role in the horse’s immune response and the study demonstrates how they can be tested using a whole blood culture using different stimulants, providing a useful tool in understanding equine health conditions.

Objective of the Research

  • The primary goal of this study was to develop an efficient, inexpensive, and reproducible method to measure TNF-α and IL-1Ra levels in equine whole blood cultures. These compounds are vital markers in the horse’s immune response.

Methodology

  • The researchers excited the blood samples with various substances: lipopolysaccharide (LPS), PCPwL (a combination of phytohemagglutinin E, concanavalin A, and pokeweed mitogen), or equine recombinant TNF-α (erTNF-α).
  • The concentrations of TNF-α and IL-1Ra in these culture samples were then measured using enzyme-linked immunosorbent assays (ELISA).
  • The study also investigated the release kinetics—how quickly and in what amount these compounds emerged—for TNF-α and IL-1Ra in the samples were studied over various time frames.

Results

  • Both TNF-α and IL-1Ra were optimally detected in blood cultures that contained a 20% dilution.
  • The release of TNF-α and IL-1Ra was time-dependent, but their release patterns differed. PCPwL caused constant enhancement in TNF-α and IL-1Ra release over 24-48 hours. In contrast, LPS-triggered TNF-α peaked between 8-12 hours and decreased afterwards, while IL-1Ra levels increased between 12-24 hours and continued to accumulate over 48 hours.
  • Introduction of equine recombinant TNF-α also stimulated IL-1Ra release in the blood samples.

Conclusion

  • This study showed that both PCPwL and LPS succeeded in stimulating time-dependent production of TNF-α and IL-1Ra in whole blood cultures. This suggests that whole blood cultures could be a useful tool for assessing the release of various cytokines, crucial components of the immune system, to better understand equine health and diseases.

Cite This Article

APA
Rütten S, Schusser GF, Abraham G, Schrödl W. (2016). Release kinetics of tumor necrosis factor-α and interleukin-1 receptor antagonist in the equine whole blood. BMC Vet Res, 12(1), 117. https://doi.org/10.1186/s12917-016-0742-4

Publication

BMC veterinary research
ISSN: 1746-6148
NlmUniqueID: 101249759
Country: England
Language: English
Volume: 12
Issue: 1
Pages: 117
PII: 117

Researcher Affiliations

Rütten, Simon
  • Institute of Pharmacology, Pharmacy and Toxicology, Faculty of Veterinary Medicine, Leipzig University, An den Tierkliniken 15, 04103, Leipzig, Germany.
Schusser, Gerald F
  • Department of Large Animal Medicine, Faculty of Veterinary Medicine, Leipzig University, An den Tierkliniken 11, 04103, Leipzig, Germany.
Abraham, Getu
  • Institute of Pharmacology, Pharmacy and Toxicology, Faculty of Veterinary Medicine, Leipzig University, An den Tierkliniken 15, 04103, Leipzig, Germany. gabraham@rz.uni-leipzig.de.
Schrödl, Wieland
  • Institute of Bacteriology and Mycology, Faculty of Veterinary Medicine, Leipzig University, An den Tierkliniken 29, 04103, Leipzig, Germany.

MeSH Terms

  • Animals
  • Blood Chemical Analysis / veterinary
  • Enzyme-Linked Immunosorbent Assay
  • Horses
  • In Vitro Techniques
  • Inflammation / veterinary
  • Interleukin 1 Receptor Antagonist Protein / blood
  • Interleukin 1 Receptor Antagonist Protein / metabolism
  • Leukocytes, Mononuclear / drug effects
  • Lipopolysaccharides / pharmacology
  • Tumor Necrosis Factor-alpha / blood
  • Tumor Necrosis Factor-alpha / metabolism

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Citations

This article has been cited 2 times.
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  2. Urayama S, Muko R, Muranaka M, Mita H, Ohta M, Matsuda H, Tanaka A. Differential effects of flunixin meglumine and meloxicam on TNF- α production in LPS-stimulated equine neutrophils in vitro. Vet Anim Sci 2025 Dec;30:100513.
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