Effect of immunosuppressive drugs on cytokine production in canine whole blood stimulated with lipopolysaccharide or a combination of ionomycin and phorbol 12‐myristate 13‐acetate

Abstract

A pharmacodynamic assay has been previously developed to monitor ciclosporin treatment in dogs by assessing inhibition of cytokine transcription after whole blood stimulation with 12‐myristate 13‐1 acetate and ionomycin ( PMA /I). In this study, whole blood stimulation with either PMA /I or lipopolysaccharide ( LPS ) was used to assess the effect of multiple drugs (azathioprine, ciclosporin, mycophenolate, leflunomide and prednisone) after a 7‐day treatment course on production of cytokines measured with a multiplex assay in healthy dogs ( n = 4 for each treatment). Interleukin‐10 ( IL ‐10), interferon gamma ( IFN γ ) and tumour necrosis factor alpha ( TNF α ) were significantly activated by PMA /I stimulation and IL ‐6, IL ‐10 and TNF α by LPS stimulation, in the absence of immunosuppressive drugs. After ciclosporin treatment, IL ‐10, IFN γ and TNF α production was significantly reduced after stimulation with PMA /I compared to pre‐treatment. After prednisone treatment, TNF α production was significantly reduced after stimulation with PMA /I or LPS compared to pre‐treatment. No significant change was observed after treatment with azathioprine, leflunomide or mycophenolate. This methodology may be useful to monitor dogs not only treated with ciclosporin, but also with prednisone or a combination of both. Further studies are needed to assess the use of this assay in a clinical setting.

Tissue‐resident mucosal‐associated invariant T (MAIT) cells in the human kidney represent a functionally distinct subset

“…This may be a consequence of more frequent infections in these patients. Furthermore, post‐transplantation, we found that a decreased percentage of circulating MAIT cells was producing TNFα, potentially due to the use of immunosuppressive agents [59]. Nevertheless, in the current study, there was no evidence to suggest that a decreased number or the impaired function of circulating MAIT cells are responsible for the frequent infections observed in RTRs both before and after transplantation.…”

Development of monoclonal antibodies for quantification of bovine tumor necrosis factor-α

“…In plasma from whole-blood stimulation, however, LPS induced a much higher concentration of TNF-α, and PMA/ionomycin-stimulated samples showed low to no detectable TNF-α (Figure 2D). Dandrieux et al (2019) observed similar results after stimulation of canine whole blood with lower TNF-α concentrations in response to PMA/ionomycin compared with LPS stimulation. The discrepancy between the response to PMA/ionomycin in PBMC and in whole blood can be associated with the difference in cell composition in the 2 samples.…”

Methylprednisolone Induces Extracellular Trap Formation and Enhances Bactericidal Effect of Canine Neutrophils

“…Significant changes in cytokine expression were also shown in dogs, especially for TNF α, treated with either high doses of ciclosporin or a standard immunosuppressive dosage of prednisolone. TNF α concentrations significantly decreased [ 104 ]. This effect was also described in a canine endotoxemia model.…”