Treatment of low doses curcumin could modulate Th17/Treg balance specifically on CD4+ T cell cultures of systemic lupus erythematosus patients

Kusworini Handono, Mirza Zaka Pratama, Agustina Tri Endharti, Handono Kalim, Kusworini Handono, Mirza Zaka Pratama, Agustina Tri Endharti, Handono Kalim

Abstract

Introduction: The balance between T helper 17 (Th17) and regulatory T cells (Treg) is a new paradigm in the pathogenesis of systemic lupus erythematosus (SLE). Currently, there are no drugs that able to modulate Th17/Treg balance specifically in SLE. Curcumin is a bioactive agent that has a specific action against hyperproliferative cells. However, its role in modulating Th17/Treg balance in SLE is still unknown. This research aimed to investigate the role of curcumin in modulating Th17/Treg balance on CD4+ T cell cultures of SLE patients.

Material and methods: CD4+ T cells from SLE 6 untreated patients and 6 healthy subjects were collected, stimulated with Th17 differentiating factors, and curcumin 0.1 and 1 µg/ml was added on cultures. After 72 hours incubation, cells were harvested and measured for Th17 and Treg percentages using flow cytometry and interleukin-17A (IL-17A) and transforming growth factor-β1 (TGF-β1) levels using ELISA.

Results: Administration of low doses of curcumin (0.1 and 1 µg/ml) could decrease Th17 percentages (p = 0.000 and p = 0.000 compared to control), reduce IL-17A productions (p = 0.000 and p = 0.000 compared to control), increase Treg percentages (p = 0.001 and p = 0.000 compared to control), and increase TGF-β1 productions (p = 0.001 and p = 0.000 compared to control) on CD4+ T cells of SLE patients. Interestingly, these effects were not reproduced on CD4+ T cells cultures of healthy subjects.

Conclusions: These data suggest that curcumin can modulate Th17/Treg balance specifically on CD4+ T cells of SLE patients without affecting healthy subjects.

Keywords: Th17/Treg balance; curcumin; systemic lupus erythematosus.

Figures

Fig. 1
Fig. 1
Comparison of Th17 and Treg profiles on CD4+ T cells cultures of healthy subjects and SLE patients before curcumin treatments. A) Representative dot plots that illustrated the expression of CD4 and IL-17A. B) Histogram of Th17 percentages expressing CD4+ IL-17A+. C) Representative dot plots that illustrated the expression of CD25 and FoxP3 which had been gated for CD4 before. D) Histogram of Treg percentages expressing CD4+ CD25+ FoxP3+. E) IL-17A levels on culture supernatant. F) TGF-β1 levels on culture supernatant. Data are presented as the mean ± standard deviation
Fig. 2
Fig. 2
Effect of curcumin treatments on apoptosis of CD4+ T cells. A) Representative dot plots that illustrated expression of Annexin V and PI. B) Histogram of viable cells, early apoptotic cells, and late apoptotic cells on each group (0, 0.1, 1, 10, and 100 µg/ml of curcumin). Data are presented as the mean ± standard deviation
Fig. 3
Fig. 3
Effect of curcumin treatments on in vitro differentiated Th17 from CD4+ T cells cultures. A) Representative dot plots that illustrated the expression of CD4 and IL-17A of SLE patients. B) Histogram of Th17 percentages expressing CD4+ IL-17A+ from CD4+ T cells cultures of SLE patients. C) Representative dot plots that illustrated the expression of CD4 and IL-17A of healthy subjects. D) Histogram of Th17 percentage expressing CD4+ IL-17A+ from CD4+ T cells cultures of healthy subjects. Data are presented as the mean ± standard deviation
Fig. 4
Fig. 4
Effect of curcumin treatments on in vitro differentiated Treg from CD4+ T cells cultures. A) Representative dot plots that illustrated the expression of CD25 and FoxP3 which had been gated for CD4 before of SLE patients. B) Histogram of Treg percentages expressing CD4+ CD25+ FoxP3+ from CD4+ T cells cultures of SLE patients. C) Representative dot plots that illustrated the expression of CD25 and FoxP3 which had been gated for CD4 before of healthy subjects. D) Histogram of Treg percentages expressing CD4+ CD25+ FoxP3+ from CD4+ T cells cultures of healthy subjects. Data are presented as the mean ± standard deviation
Fig. 5
Fig. 5
Effect of curcumin treatments on IL-17A and TGF-β1 productions. A) Histogram of IL-17A levels from culture supernatant of SLE patients. B) Histogram of TGF-β1 levels from culture supernatant of SLE patients. C) Histogram of IL-17A levels from culture supernatant of healthy subjects. D) Histogram of TGF-β1 levels from culture supernatant of healthy subjects. Data are presented as the mean ± standard deviation

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