Influences of age and sex on leukocytes of healthy horses and their ex vivo cytokine release.
Abstract: Leukocytes and their functional capacities are used extensively as biomarkers in immunological research. Commonly employed indicators concerning leukocytes are as follows: number, composition in blood, response to discrete stimuli, cytokine release, and morphometric characteristics. In order to employ leukocytes as biomarkers for disease and therapeutic monitoring, physiological variations and influencing factors on the parameters measured have to be considered. The aim of this report was to describe the ranges of selected leukocyte parameters in a sample of healthy horses and to analyse whether age, sex, breed, and sampling time point (time of day) influence peripheral blood leukocyte composition, cell morphology and release of cytokines ex vivo. Flow cytometric comparative characterisation of cell size and complexity in 24 healthy horses revealed significant variance. Similarly, basal release of selected cytokines by blood mononuclear cells also showed high variability [TNFα (65-16,624pg/ml), IFNγ (4-80U/ml), IL-4 (0-5069pg/ml), IL-10 (49-1862pg/ml), and IL-17 (4-1244U/ml)]. Each animal's age influenced leukocyte composition, cell morphology and cytokine release (TNFα, IL-4, IL-10) ex vivo. Geldings showed smaller monocytes and higher spontaneous production of IL-10 when compared to the mares included. The stimulation to spontaneous release ratios of TNFα, IL-4 and IL-17 differed in Warmblood and Thoroughbred types. Sampling time influenced leukocyte composition and cell morphology. In summary, many animal factors - age being the dominant one - should be considered for studies involving the analysis of equine leukocytes. In addition, high inter-individual variances argue for individual baseline measurements.
Copyright © 2015 Elsevier B.V. All rights reserved.
Publication Date: 2015-03-06 PubMed ID: 25782350DOI: 10.1016/j.vetimm.2015.02.011Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research investigated the impact of various factors on various features of horse leukocytes, such as their number, composition, responsiveness to stimuli, cytokine release, and morphometric characteristics. The study found that the age of the horse, in particular, significantly influences these parameters.
Overview and Objective of the Study
- The study seeks to establish the influence of various factors like age, sex, breed, and sampling time point on leukocyte parameters in horses. Leukocytes are white blood cells that play an integral part in the body’s immune system to fight infections.
- The aim is to understand these influences to better employ leukocytes as biomarkers for monitoring diseases and the effects of therapy.
Methodology and Findings
- 24 healthy horses underwent flow cytometric measurements of cell size and complexity, unveiling significant variations.
- Basal release levels of certain cytokines showed high variability as well. Cytokines are proteins that help in cell signaling and guide the development and response of immune cells.
- Individual horse’s age affected its leukocyte profile. Older horses showed different leukocyte composition, cell morphology, and cytokine release levels compared to younger ones.
Impacts of Sex and Breed
- The team found gender differences in leukocytes. Geldings (castrated males) had smaller monocytes and higher spontaneous production of a protein called IL-10 than the mares in the study.
- Breed also affected leukocytes with distinct differences in cytokine release between Warmblood and Thoroughbred types.
Time-Based Variations
- Sampling time, or the time of day when samples were collected, also influenced observations. This factor resulted in different leukocyte composition and cell morphology, further confirming the need to consider temporal aspects in such studies.
Conclusions and Implications
- In conclusion, many factors, particularly age, can affect the analysis of equine leukocytes. As such, these factors need to be taken into account when conducting such immune-based studies.
- A horse’s individuality also plays a significant role, with the study arguing for the need for individual baseline measurements due to high inter-individual variations.
Cite This Article
APA
Schnabel CL, Steinig P, Schuberth HJ, Koy M, Wagner B, Wittig B, Juhls C, Willenbrock S, Murua Escobar H, Jaehnig P, Feige K, Cavalleri JM.
(2015).
Influences of age and sex on leukocytes of healthy horses and their ex vivo cytokine release.
Vet Immunol Immunopathol, 165(1-2), 64-74.
https://doi.org/10.1016/j.vetimm.2015.02.011 Publication
Researcher Affiliations
- University of Veterinary Medicine Hannover, Foundation, Clinic for Horses, Buenteweg 9, 30559 Hannover, Germany. Electronic address: Christiane.Schnabel@tiho-hannover.de.
- University of Veterinary Medicine Hannover, Foundation, Clinic for Horses, Buenteweg 9, 30559 Hannover, Germany.
- University of Veterinary Medicine Hannover, Foundation, Immunology Unit, Bischofsholer Damm 15, 30173 Hannover, Germany.
- University of Veterinary Medicine Hannover, Foundation, Immunology Unit, Bischofsholer Damm 15, 30173 Hannover, Germany.
- Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University College of Veterinary Medicine, 240 Farrier Rd, Ithaca, NY 14853, USA.
- Foundation Institute Molecular Biology and Bioinformatics, Freie Universitaet Berlin, Berlin, Germany.
- Mologen AG, Fabeckstrasse 30, 14195 Berlin, Germany.
- University of Veterinary Medicine Hannover, Foundation, Small Animal Clinic, Buenteweg 9, 30559 Hannover, Germany.
- University of Veterinary Medicine Hannover, Foundation, Small Animal Clinic, Buenteweg 9, 30559 Hannover, Germany; University of Rostock, Division of Medicine, Clinic III, Haematology, Oncology and Palliative Medicine, 18057 Rostock, Germany.
- pj statistics, Niedstrasse. 16, 12159 Berlin, Germany.
- University of Veterinary Medicine Hannover, Foundation, Clinic for Horses, Buenteweg 9, 30559 Hannover, Germany.
- University of Veterinary Medicine Hannover, Foundation, Clinic for Horses, Buenteweg 9, 30559 Hannover, Germany.
MeSH Terms
- Age Factors
- Animals
- Cytokines / blood
- Cytokines / physiology
- Female
- Flow Cytometry / veterinary
- Horses / immunology
- Horses / physiology
- Interferon-gamma / blood
- Interferon-gamma / physiology
- Interleukin-10 / blood
- Interleukin-10 / physiology
- Interleukin-17 / blood
- Interleukin-17 / physiology
- Interleukin-4 / blood
- Interleukin-4 / physiology
- Leukocytes / metabolism
- Leukocytes / physiology
- Male
- Sex Factors
- Tumor Necrosis Factor-alpha / blood
- Tumor Necrosis Factor-alpha / physiology
Citations
This article has been cited 9 times.- DeNotta S, McFarlane D. Immunosenescence and inflammaging in the aged horse. Immun Ageing 2023 Jan 6;20(1):2.
- Dabravolski SA, Nikiforov NG, Zhuravlev AD, Orekhov NA, Grechko AV, Orekhov AN. Role of the mtDNA Mutations and Mitophagy in Inflammaging. Int J Mol Sci 2022 Jan 25;23(3).
- Hellman S, Hjertner B, Morein B, Fossum C. The adjuvant G3 promotes a Th1 polarizing innate immune response in equine PBMC. Vet Res 2018 Oct 22;49(1):108.
- Parkinson NJ, Buechner-Maxwell VA, Witonsky SG, Pleasant RS, Werre SR, Ahmed SA. Characterization of basal and lipopolysaccharide-induced microRNA expression in equine peripheral blood mononuclear cells using Next-Generation Sequencing. PLoS One 2017;12(5):e0177664.
- Geburek F, Gaus M, van Schie HT, Rohn K, Stadler PM. Effect of intralesional platelet-rich plasma (PRP) treatment on clinical and ultrasonographic parameters in equine naturally occurring superficial digital flexor tendinopathies - a randomized prospective controlled clinical trial. BMC Vet Res 2016 Sep 7;12(1):191.
- Schnabel CL, Steinig P, Koy M, Schuberth HJ, Juhls C, Oswald D, Wittig B, Willenbrock S, Murua Escobar H, Pfarrer C, Wagner B, Jaehnig P, Moritz A, Feige K, Cavalleri JM. Immune response of healthy horses to DNA constructs formulated with a cationic lipid transfection reagent. BMC Vet Res 2015 Jun 23;11:140.
- Aragona F, Rizzo M, Giudice E, Fazio F, Costa A, Di Bella B, De Caro S, Arfuso F, Briglia M, Piccione G, Giannetto C. Circadian Oscillation of Leukocyte Subpopulations and Inflammatory Cytokines over a 24-H Period in Horses. Vet Sci 2025 Apr 20;12(4).
- Hartwig S, Burron S, Richards T, Rankovic A, Ma DWL, Pearson W, Ellis J, Trevizan L, Seymour DJ, Shoveller AK. The effect of dietary camelina, flaxseed, and canola oil supplementation on skin fatty acid profile and immune and inflammatory responses in healthy adult horses. J Anim Sci 2025 Jan 4;103.
- Harari S, Deretz S, Dumont Saint Priest B, Richard E, Ricard A. Comparison of blood parameters in two genetically different groups of horses for functional longevity in show jumping. Front Genet 2024;15:1455790.
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