Effects of cyclosporine and dexamethasone on canine T cell expression of interleukin-2 and interferon-gamma.
Vet Immunol Immunopathol
; 216: 109892, 2019 Oct.
Article
em En
| MEDLINE
| ID: mdl-31446206
ABSTRACT
Cyclosporine and glucocorticoids are powerful immunosuppressive agents used to treat many inflammatory diseases in dogs. Cyclosporine inhibits calcineurin-dependent pathways of T cell activation and resultant T cell cytokine production, and glucocorticoids directly inhibit genes coding for cytokines. Little work has been done comparing the effects of these agents on T cell cytokine production in dogs. Our study measured T cell interleukin-2 (IL-2) and interferon-gamma (IFN-γ) production using flow cytometry and T cell IL-2 and IFN-γ gene expression using quantitative reverse transcription polymerase chain reaction (qRT-PCR) in activated canine T cells incubated with cyclosporine and dexamethasone in vitro. For flow cytometric assays, diluted whole blood was cultured for 7â¯h in the presence of cyclosporine (10, 100, 500, and 1000â¯ng/mL) or dexamethasone (10â¯ng/mL, 100â¯ng/mL, 1⯵g/mL, and 10⯵g/mL). For qRT-PCR, whole blood was cultured for 5â¯h with the same drugs at the same concentrations, and RNA was then extracted from leukocytes. Flow cytometry and qRT-PCR both demonstrated inhibition of IL-2 and IFN-γ that was concentration-dependent in response to cyclosporine, and was more variable for dexamethasone. Quantitative RT-PCR but not flow cytometry documented significant reduction of IL-2 expression after dexamethasone treatment, while both methods showed concentration-dependent suppression of IFN-γ. Quantitative RT-PCR also revealed additional cytokine suppression at higher cyclosporine concentrations, an effect not found using flow cytometry, and may therefore be the preferred method for cytokine determination in dogs. Suppression of IL-2 and IFN-γ in activated T cells may have potential as an indicator of the efficacy of cyclosporine and glucocorticoids in suppressing canine T cell function in vivo, and may therefore be of value for characterizing the immunosuppression induced by these drugs in clinical patients.
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Bases de dados:
MEDLINE
Assunto principal:
Dexametasona
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Linfócitos T
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Regulação da Expressão Gênica
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Interferon gama
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Interleucina-2
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Ciclosporina
Limite:
Animals
Idioma:
En
Revista:
Vet Immunol Immunopathol
Ano de publicação:
2019
Tipo de documento:
Article