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PloS one2012; 7(9); e44606; doi: 10.1371/journal.pone.0044606

Neutrophils are not less sensitive than other blood leukocytes to the genomic effects of glucocorticoids.

Abstract: Neutrophils are generally considered less responsive to glucocorticoids compared to other inflammatory cells. The reported increase in human neutrophil survival mediated by these drugs partly supports this assertion. However, it was recently shown that dexamethasone exerts potent anti-inflammatory effects in equine peripheral blood neutrophils. Few comparative studies of glucocorticoid effects in neutrophils and other leukocytes have been reported and a relative insensitivity of neutrophils to these drugs could not be ruled out. Objective: We assessed glucocorticoid-responsiveness in equine and human peripheral blood neutrophils and neutrophil-depleted leukocytes. Methods: Blood neutrophils and neutrophil-depleted leukocytes were isolated from 6 healthy horses and 4 human healthy subjects. Cells were incubated for 5 h with or without LPS (100 ng/mL) alone or combined with hydrocortisone, prednisolone or dexamethasone (10(-8) M and 10(-6) M). IL-1β, TNF-α, IL-8, glutamine synthetase and GR-α mRNA expression was quantified by qPCR. Equine neutrophils were also incubated for 20 h with or without the three glucocorticoids and cell survival was assessed by flow cytometry and light microscopy on cytospin preparations. Results: We found that glucocorticoids down-regulated LPS-induced pro-inflammatory mRNA expression in both cell populations and species. These drugs also significantly increased glutamine synthetase gene expression in both equine cell populations. The magnitude of glucocorticoid response between cell populations was generally similar in both species. We also showed that dexamethasone had a comparable inhibitory effect on pro-inflammatory gene expression in both human and equine neutrophils. As reported in other species, glucocorticoids significantly increase the survival in equine neutrophils. Conclusions: Glucocorticoids exert genomic effects of similar magnitude on neutrophils and on other blood leukocytes. We speculate that the poor response to glucocorticoids observed in some chronic neutrophilic diseases such as severe asthma or COPD is not explained by a relative lack of inhibition of these drugs on pro-inflammatory cytokines expression in neutrophils.
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Publication Date: 2012-09-12 PubMed ID: 22984532PubMed Central: PMC3440353DOI: 10.1371/journal.pone.0044606Google Scholar: Lookup
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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 study investigates the effects of glucocorticoids – a type of steroid hormone – on different types of white blood cells, specifically neutrophils. The researchers conclude that neutrophils are not less sensitive to the effects of glucocorticoids compared to other white blood cells, contrary to the common belief.

Objective and Methodology of the Research

  • The study aimed to assess the responsiveness of neutrophils – a type of leukocyte (white blood cell) – to glucocorticoids in equine and human blood.
  • The researchers isolated blood neutrophils and neutrophil-depleted leukocytes from six healthy horses and four humans.
  • The isolated cells were incubated for five hours with or without LPS (a molecule found in bacteria that triggers inflammation) either on its own or in combination with one of three glucocorticoids: hydrocortisone, prednisolone, or dexamethasone. The researchers also carried out a longer, 20-hour incubation with the glucocorticoids for equine neutrophils.
  • The researchers then measured the expression of several genes associated with inflammation and glucocorticoid response using quantitative PCR; a method for amplifying and measuring specific DNA sequences.

Results of the Research

  • The research showed that glucocorticoids down-regulated the expression of genes associated with pro-inflammation in both the neutrophils and the other leukocytes in both humans and horses. This means that the glucocorticoids were able to reduce the inflammatory response in these cells.
  • Moreover, glucocorticoids robustly increased the expression of the glutamine synthetase gene in both types of equine cells. As glutamine synthetase plays a key role in certain pathways related to cellular energy and the immune system, this suggests another way that glucocorticoids may be affecting these cells.
  • The researchers also found that the glucocorticoid dexamethasone was similarly effective at inhibiting the expression of pro-inflammatory genes in both human and equine neutrophils.
  • As observed in other species, glucocorticoids significantly increased the survival rate of equine neutrophils.

Conclusions of the Research

  • The researchers concluded that glucocorticoids have similar effects on neutrophils and other leukocytes, contradicting the current belief that neutrophils are less responsive to these drugs.
  • They suggest that the ineffective response of some chronic neutrophilic diseases like severe asthma or COPD to glucocorticoids is not due to an inherent insensitivity of neutrophils to these drugs.

Cite This Article

APA
Hirsch G, Lavoie-Lamoureux A, Beauchamp G, Lavoie JP. (2012). Neutrophils are not less sensitive than other blood leukocytes to the genomic effects of glucocorticoids. PLoS One, 7(9), e44606. https://doi.org/10.1371/journal.pone.0044606

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 7
Issue: 9
Pages: e44606
PII: e44606

Researcher Affiliations

Hirsch, Gaelle
  • Department of Clinical Sciences, Faculty of Veterinary Medicine, Université de Montréal, Saint-Hyacinthe, Q, Canada.
Lavoie-Lamoureux, Anouk
    Beauchamp, Guy
      Lavoie, Jean-Pierre

        MeSH Terms

        • Adult
        • Animals
        • Anti-Inflammatory Agents / pharmacology
        • Cell Separation
        • Cell Survival
        • Cells, Cultured
        • Dose-Response Relationship, Drug
        • Down-Regulation
        • Female
        • Flow Cytometry / methods
        • Gene Expression Regulation / drug effects
        • Glucocorticoids / metabolism
        • Glucocorticoids / pharmacology
        • Horses
        • Humans
        • Leukocytes / drug effects
        • Lipopolysaccharides / metabolism
        • Neutrophils / drug effects
        • RNA, Messenger / metabolism

        Grant Funding

        • MOP-94848 / Canadian Institutes of Health Research

        Conflict of Interest Statement

        The authors have declared that no competing interests exist.

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