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Lack of in vitro effect of aglepristone on IFN-γ and IL-4 production by resting and mitogen-activated T cells of luteal bitches.

Jurka P, Szulc-Dąbrowska L, Borkowska J, Winnicka A - BMC Vet. Res. (2013)

Bottom Line: Our results showed no statistically significant differences in the percentage of IFN-γ and IL-4-synthesizing CD4+ or CD8+ resting T cells between untreated and aglepristone-treated cells at 24 and 48 hours post treatment.Presented results indicate that administration of aglepristone for 48 hours has no influence on IFN-γ and IL-4 synthesis by resting and mitogen-activated T cells isolated from diestral bitches.We conclude that antiprogestins may differentially affect T cell function depending on the animal species in which they are applied.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Small Animal Diseases with Clinic, Laboratory of Small Animal Reproduction, Warsaw, Poland. piotr_jurka@sggw.pl.

ABSTRACT

Background: Aglepristone (RU534) is an antiprogestin used for pregnancy termination, parturition induction and conservative pyometra treatment in bitches. Its molecular structure is similar to mifepristone, an antiprogestin used in human medicine. Mifepristone has been shown to suppress proliferation and cytokine production by T cells, whereas the effect of aglepristone on T cell function remains elusive. The purpose of this project was to investigate the in vitro influence of RU534 on IFN-γ and IL-4 synthesis by peripheral blood T cells isolated from healthy bitches (N = 16) in luteal phase. The peripheral blood mononuclear cells (PBMCs) were incubated with three different dosages of aglepristone, or dimethyl sulfoxide (DMSO), with or without mitogen. The production of cytokines by resting or mitogen-activated T cells was determined by intercellular staining and flow cytometry analysis or ELISA assay, respectively.

Results: Our results showed no statistically significant differences in the percentage of IFN-γ and IL-4-synthesizing CD4+ or CD8+ resting T cells between untreated and aglepristone-treated cells at 24 and 48 hours post treatment. Moreover, mitogen-activated PBMCs treated with RU534 displayed similar concentration of IFN-γ and IL-4 in culture supernatants to those observed in mitogen-activated DMSO-treated PBMCs. Presented results indicate that administration of aglepristone for 48 hours has no influence on IFN-γ and IL-4 synthesis by resting and mitogen-activated T cells isolated from diestral bitches.

Conclusions: We conclude that antiprogestins may differentially affect T cell function depending on the animal species in which they are applied.

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Related in: MedlinePlus

The percentage of IFN-γ-synthesizing T cell subsets (A) and the MFI of IFN-γ-positive T cell subsets (B) treated for 24 and 48 h with 30, 300 and 3000 ng/ml of aglepristone. All data are presented relative to control values (line). Each point represents an individual dog (n = 7–9), with the bar indicating the mean. *P < 0.05, ** P < 0.01.
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Figure 2: The percentage of IFN-γ-synthesizing T cell subsets (A) and the MFI of IFN-γ-positive T cell subsets (B) treated for 24 and 48 h with 30, 300 and 3000 ng/ml of aglepristone. All data are presented relative to control values (line). Each point represents an individual dog (n = 7–9), with the bar indicating the mean. *P < 0.05, ** P < 0.01.

Mentions: Representative dot blots showing synthesis of IFN-γ by CD4+ and CD8+ T cells isolated from diestral bitches and in vitro treated with DMSO (control) or 30, 300 and 3000 ng/ml of aglepristone, are presented on Figure 1. The percentage of CD4+ T cells-synthesizing IFN-γ ranged from 6.0% to 18.1% and 4.7% to 18.6% in control cells, 5.2% to 27.7% and 4.2% to 24.0% in cells treated with 30 ng/ml of aglepristone, 4.3% to 22.1% and 6.9% to 20.5% in cells treated with 300 ng/ml of aglepristone, 5.2% to 26.3% and 4.1% to 20.2% in cells treated with 3000 ng/ml of aglepristone, at 24 and 48 h of incubation, respectively. Our results showed no differences in the percentage of CD4+ T cells-synthesizing IFN-γ between control and studied groups (Figure 2A). Moreover, flow cytometric analysis revealed no effect of aglepristone on MFI of IFN-γ produced by CD4+ T cells (Figure 2B). The percentage of CD8+ T cells-synthesizing IFN-γ in control and aglepristone-treated groups ranged from 15% to around 60% at 24 and 48 h of incubation. Similarly to CD4+ T cells, CD8+ T cells synthesized IFN-γ at similar levels in the control and aglepristone-treated groups (Figure 2).


Lack of in vitro effect of aglepristone on IFN-γ and IL-4 production by resting and mitogen-activated T cells of luteal bitches.

Jurka P, Szulc-Dąbrowska L, Borkowska J, Winnicka A - BMC Vet. Res. (2013)

The percentage of IFN-γ-synthesizing T cell subsets (A) and the MFI of IFN-γ-positive T cell subsets (B) treated for 24 and 48 h with 30, 300 and 3000 ng/ml of aglepristone. All data are presented relative to control values (line). Each point represents an individual dog (n = 7–9), with the bar indicating the mean. *P < 0.05, ** P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3818567&req=5

Figure 2: The percentage of IFN-γ-synthesizing T cell subsets (A) and the MFI of IFN-γ-positive T cell subsets (B) treated for 24 and 48 h with 30, 300 and 3000 ng/ml of aglepristone. All data are presented relative to control values (line). Each point represents an individual dog (n = 7–9), with the bar indicating the mean. *P < 0.05, ** P < 0.01.
Mentions: Representative dot blots showing synthesis of IFN-γ by CD4+ and CD8+ T cells isolated from diestral bitches and in vitro treated with DMSO (control) or 30, 300 and 3000 ng/ml of aglepristone, are presented on Figure 1. The percentage of CD4+ T cells-synthesizing IFN-γ ranged from 6.0% to 18.1% and 4.7% to 18.6% in control cells, 5.2% to 27.7% and 4.2% to 24.0% in cells treated with 30 ng/ml of aglepristone, 4.3% to 22.1% and 6.9% to 20.5% in cells treated with 300 ng/ml of aglepristone, 5.2% to 26.3% and 4.1% to 20.2% in cells treated with 3000 ng/ml of aglepristone, at 24 and 48 h of incubation, respectively. Our results showed no differences in the percentage of CD4+ T cells-synthesizing IFN-γ between control and studied groups (Figure 2A). Moreover, flow cytometric analysis revealed no effect of aglepristone on MFI of IFN-γ produced by CD4+ T cells (Figure 2B). The percentage of CD8+ T cells-synthesizing IFN-γ in control and aglepristone-treated groups ranged from 15% to around 60% at 24 and 48 h of incubation. Similarly to CD4+ T cells, CD8+ T cells synthesized IFN-γ at similar levels in the control and aglepristone-treated groups (Figure 2).

Bottom Line: Our results showed no statistically significant differences in the percentage of IFN-γ and IL-4-synthesizing CD4+ or CD8+ resting T cells between untreated and aglepristone-treated cells at 24 and 48 hours post treatment.Presented results indicate that administration of aglepristone for 48 hours has no influence on IFN-γ and IL-4 synthesis by resting and mitogen-activated T cells isolated from diestral bitches.We conclude that antiprogestins may differentially affect T cell function depending on the animal species in which they are applied.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Small Animal Diseases with Clinic, Laboratory of Small Animal Reproduction, Warsaw, Poland. piotr_jurka@sggw.pl.

ABSTRACT

Background: Aglepristone (RU534) is an antiprogestin used for pregnancy termination, parturition induction and conservative pyometra treatment in bitches. Its molecular structure is similar to mifepristone, an antiprogestin used in human medicine. Mifepristone has been shown to suppress proliferation and cytokine production by T cells, whereas the effect of aglepristone on T cell function remains elusive. The purpose of this project was to investigate the in vitro influence of RU534 on IFN-γ and IL-4 synthesis by peripheral blood T cells isolated from healthy bitches (N = 16) in luteal phase. The peripheral blood mononuclear cells (PBMCs) were incubated with three different dosages of aglepristone, or dimethyl sulfoxide (DMSO), with or without mitogen. The production of cytokines by resting or mitogen-activated T cells was determined by intercellular staining and flow cytometry analysis or ELISA assay, respectively.

Results: Our results showed no statistically significant differences in the percentage of IFN-γ and IL-4-synthesizing CD4+ or CD8+ resting T cells between untreated and aglepristone-treated cells at 24 and 48 hours post treatment. Moreover, mitogen-activated PBMCs treated with RU534 displayed similar concentration of IFN-γ and IL-4 in culture supernatants to those observed in mitogen-activated DMSO-treated PBMCs. Presented results indicate that administration of aglepristone for 48 hours has no influence on IFN-γ and IL-4 synthesis by resting and mitogen-activated T cells isolated from diestral bitches.

Conclusions: We conclude that antiprogestins may differentially affect T cell function depending on the animal species in which they are applied.

Show MeSH
Related in: MedlinePlus