Effect of hydrogen peroxide on the oxidative burst of neutrophils in pigs and ruminants

Background and Aim: Neutrophils represent between 20% and 75% of white blood cells in animals and play a key role in an effective immune response. The generation of reactive oxygen species (ROS) is commonly referred to as an oxidative burst and is crucial under healthy and disease conditions. Intere...

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Main Authors: Francesco Mosca, Abigail R. Trachtman, Jasmine Hattab, Giuseppe Marruchella, Pietro G. Tiscar
Format: Article
Language:English
Published: Veterinary World 2020-09-01
Series:Veterinary World
Subjects:
Online Access:http://www.veterinaryworld.org/Vol.13/September-2020/26.pdf
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author Francesco Mosca
Abigail R. Trachtman
Jasmine Hattab
Giuseppe Marruchella
Pietro G. Tiscar
author_facet Francesco Mosca
Abigail R. Trachtman
Jasmine Hattab
Giuseppe Marruchella
Pietro G. Tiscar
author_sort Francesco Mosca
collection DOAJ
description Background and Aim: Neutrophils represent between 20% and 75% of white blood cells in animals and play a key role in an effective immune response. The generation of reactive oxygen species (ROS) is commonly referred to as an oxidative burst and is crucial under healthy and disease conditions. Interestingly, ROS are emerging as regulators of several neutrophil functions, including their oxidative burst. The present study aimed to investigate the effect of hydrogen peroxide on the oxidative burst of neutrophils, collected from domestic animal species (namely, pig, cattle, and sheep), and exposed to different stimuli. Materials and Methods: A total of 65 slaughtered animals were included in the present study: Twenty-two pigs, 21 cattle, and 22 sheep. Blood samples were collected at bleeding and neutrophils were then purified using ad hoc developed and species-specific protocols. Neutrophils were treated with hydrogen peroxide at micromolar-to-millimolar concentrations, alone, or combined with other stimuli (i.e., opsonized yeasts, and phorbol 12-myristate 13-acetate). The generation of ROS was evaluated using a luminol-derived chemiluminescence (CL) assay. For each animal species, data were aggregated and reported as mean area under curve±standard deviation. Finally, data were statistically analyzed by one-way ANOVA, followed by Tukey's post hoc test. Results: Exposure of bovine and ovine neutrophils to hydrogen peroxide alone resulted in a dose-dependent enhancement of the CL response, which was significantly stronger at its highest concentration and proved particularly prominent in sheep. Opsonized yeasts and phorbol 12-myristate 13-acetate both proved capable of stimulating the generation of ROS in all animal species under study. Hydrogen peroxide negatively modulated the oxidative burst of neutrophils after exposure to those stimuli, observed response patterns varying between pigs and ruminants. Porcine neutrophils, pre-exposed to micromolar concentrations of hydrogen peroxide, showed a decreased CL response only to opsonized yeasts. Conversely, pre-exposure to hydrogen peroxide reduced the CL response of ruminant neutrophils both to yeasts and phorbol 12-myristate 13-acetate, the effect being most prominent at 1 mM concentration. Conclusion: These results indicate that hydrogen peroxide is capable of modulating the oxidative bursts of neutrophils in a species-specific and dose-dependent manner, substantial differences existing between pigs and ruminants. Further investigation is required to fully comprehend such modulation, which is crucial for the proper management of the generation of ROS under healthy and disease conditions.
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spelling doaj.art-2e859df42acd491b8a3bfda5110879812022-12-21T18:28:58ZengVeterinary WorldVeterinary World0972-89882231-09162020-09-011391934193910.14202/vetworld.2020.1934-1939Effect of hydrogen peroxide on the oxidative burst of neutrophils in pigs and ruminantsFrancesco Mosca0https://orcid.org/0000-0001-8188-6306Abigail R. Trachtman1https://orcid.org/0000-0003-2802-5122Jasmine Hattab2https://orcid.org/0000-0003-3711-4450Giuseppe Marruchella3https://orcid.org/0000-0003-3176-3847Pietro G. Tiscar4https://orcid.org/0000-0002-2408-0821Department of Veterinary Medicine, University of Teramo, Loc. Piano d'Accio, 64100, Teramo, Italy.Department of Veterinary Medicine, University of Teramo, Loc. Piano d'Accio, 64100, Teramo, Italy.Department of Veterinary Medicine, University of Teramo, Loc. Piano d'Accio, 64100, Teramo, Italy.Department of Veterinary Medicine, University of Teramo, Loc. Piano d'Accio, 64100, Teramo, Italy.Department of Veterinary Medicine, University of Teramo, Loc. Piano d'Accio, 64100, Teramo, Italy.Background and Aim: Neutrophils represent between 20% and 75% of white blood cells in animals and play a key role in an effective immune response. The generation of reactive oxygen species (ROS) is commonly referred to as an oxidative burst and is crucial under healthy and disease conditions. Interestingly, ROS are emerging as regulators of several neutrophil functions, including their oxidative burst. The present study aimed to investigate the effect of hydrogen peroxide on the oxidative burst of neutrophils, collected from domestic animal species (namely, pig, cattle, and sheep), and exposed to different stimuli. Materials and Methods: A total of 65 slaughtered animals were included in the present study: Twenty-two pigs, 21 cattle, and 22 sheep. Blood samples were collected at bleeding and neutrophils were then purified using ad hoc developed and species-specific protocols. Neutrophils were treated with hydrogen peroxide at micromolar-to-millimolar concentrations, alone, or combined with other stimuli (i.e., opsonized yeasts, and phorbol 12-myristate 13-acetate). The generation of ROS was evaluated using a luminol-derived chemiluminescence (CL) assay. For each animal species, data were aggregated and reported as mean area under curve±standard deviation. Finally, data were statistically analyzed by one-way ANOVA, followed by Tukey's post hoc test. Results: Exposure of bovine and ovine neutrophils to hydrogen peroxide alone resulted in a dose-dependent enhancement of the CL response, which was significantly stronger at its highest concentration and proved particularly prominent in sheep. Opsonized yeasts and phorbol 12-myristate 13-acetate both proved capable of stimulating the generation of ROS in all animal species under study. Hydrogen peroxide negatively modulated the oxidative burst of neutrophils after exposure to those stimuli, observed response patterns varying between pigs and ruminants. Porcine neutrophils, pre-exposed to micromolar concentrations of hydrogen peroxide, showed a decreased CL response only to opsonized yeasts. Conversely, pre-exposure to hydrogen peroxide reduced the CL response of ruminant neutrophils both to yeasts and phorbol 12-myristate 13-acetate, the effect being most prominent at 1 mM concentration. Conclusion: These results indicate that hydrogen peroxide is capable of modulating the oxidative bursts of neutrophils in a species-specific and dose-dependent manner, substantial differences existing between pigs and ruminants. Further investigation is required to fully comprehend such modulation, which is crucial for the proper management of the generation of ROS under healthy and disease conditions.http://www.veterinaryworld.org/Vol.13/September-2020/26.pdfhydrogen peroxideluminol-derived chemiluminescencepigpolymorphonuclear neutrophilsrespiratory burstruminants
spellingShingle Francesco Mosca
Abigail R. Trachtman
Jasmine Hattab
Giuseppe Marruchella
Pietro G. Tiscar
Effect of hydrogen peroxide on the oxidative burst of neutrophils in pigs and ruminants
Veterinary World
hydrogen peroxide
luminol-derived chemiluminescence
pig
polymorphonuclear neutrophils
respiratory burst
ruminants
title Effect of hydrogen peroxide on the oxidative burst of neutrophils in pigs and ruminants
title_full Effect of hydrogen peroxide on the oxidative burst of neutrophils in pigs and ruminants
title_fullStr Effect of hydrogen peroxide on the oxidative burst of neutrophils in pigs and ruminants
title_full_unstemmed Effect of hydrogen peroxide on the oxidative burst of neutrophils in pigs and ruminants
title_short Effect of hydrogen peroxide on the oxidative burst of neutrophils in pigs and ruminants
title_sort effect of hydrogen peroxide on the oxidative burst of neutrophils in pigs and ruminants
topic hydrogen peroxide
luminol-derived chemiluminescence
pig
polymorphonuclear neutrophils
respiratory burst
ruminants
url http://www.veterinaryworld.org/Vol.13/September-2020/26.pdf
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