CD4+ T cells에서 백개자가 IFN-$\\gamma$와 IL-4 생성에 미치는 영향 Effect of Sinapis alba L. on expression of interferon-gamma and interleukin-4 production in anti-CD3/anti-CD28-stimulated CD4(+) T cells원문보기
Objective : Sinapis alba L. (SA) is a korean traditional herbal medicine that is usually used to prevent or treat inflammatory diseases, such as respiratory infection and rheumatoid arthritis. However, the effects of SA supplementation in vitro on serum antibody levels, splenocyte and peritoneal mac...
Objective : Sinapis alba L. (SA) is a korean traditional herbal medicine that is usually used to prevent or treat inflammatory diseases, such as respiratory infection and rheumatoid arthritis. However, the effects of SA supplementation in vitro on serum antibody levels, splenocyte and peritoneal macrophage immune responses have not yet been determined. In this study, we examined the effect of SA on the production of Th1/Th2 cytokines. Methods : Splenocytes were isolated from naive C57BL/6 mice. Cells were enriched for CD4+ cell populations by first staining the cells with anti-CD4 (BD PharMingen, Calif, USA). CD4+ T cells were selected on a (CS) column, and the flow-through was collected as CD4+ T cells. Isolated cells were activated by overnight incubation on 24-well plates coated with $1{\mu}g/mL$ anti-CD3, $1{\mu}g/mL$ anti-CD28 and with SA ($100{\mu}g/mL$). Primary macrophages were collected from the peritoneal cavities of mice (8-week-old female C57BL/6). The peritoneal macrophages were washed and plated with RPMI-1640 overnight for the experiments. After 48-hours cultures, samples were centrifuged at 2000 rpm for 10 minutes, and the supernatants were stored at $-80^{\circ}C$. Mouse IL-4, IFN-$\gamma$ and TNF-$\alpha$ were quantified using ELISA kits (BioSource International, Camarillo, Calif, USA) according to the manufacturer's protocols. Results : SA at 100ug/ml decreased the generation of Th1 cytokine (IFN-$\gamma$) by 0.5-fold. However, SA has no effect on Th2 (IL-4) production. Conclusions : These results suggest that SA may play an important role in the control of T-cell-mediated autoimmunity by down-regulation of Th1 cytokine (especially IFN-$\gamma$, TNF-$\alpha$). These data may contribute to the design of new immunomodulating treatments for a group of autoimmune diseases.
Objective : Sinapis alba L. (SA) is a korean traditional herbal medicine that is usually used to prevent or treat inflammatory diseases, such as respiratory infection and rheumatoid arthritis. However, the effects of SA supplementation in vitro on serum antibody levels, splenocyte and peritoneal macrophage immune responses have not yet been determined. In this study, we examined the effect of SA on the production of Th1/Th2 cytokines. Methods : Splenocytes were isolated from naive C57BL/6 mice. Cells were enriched for CD4+ cell populations by first staining the cells with anti-CD4 (BD PharMingen, Calif, USA). CD4+ T cells were selected on a (CS) column, and the flow-through was collected as CD4+ T cells. Isolated cells were activated by overnight incubation on 24-well plates coated with $1{\mu}g/mL$ anti-CD3, $1{\mu}g/mL$ anti-CD28 and with SA ($100{\mu}g/mL$). Primary macrophages were collected from the peritoneal cavities of mice (8-week-old female C57BL/6). The peritoneal macrophages were washed and plated with RPMI-1640 overnight for the experiments. After 48-hours cultures, samples were centrifuged at 2000 rpm for 10 minutes, and the supernatants were stored at $-80^{\circ}C$. Mouse IL-4, IFN-$\gamma$ and TNF-$\alpha$ were quantified using ELISA kits (BioSource International, Camarillo, Calif, USA) according to the manufacturer's protocols. Results : SA at 100ug/ml decreased the generation of Th1 cytokine (IFN-$\gamma$) by 0.5-fold. However, SA has no effect on Th2 (IL-4) production. Conclusions : These results suggest that SA may play an important role in the control of T-cell-mediated autoimmunity by down-regulation of Th1 cytokine (especially IFN-$\gamma$, TNF-$\alpha$). These data may contribute to the design of new immunomodulating treatments for a group of autoimmune diseases.
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문제 정의
It has become apparent that, in this experiment, CD4+ T cell subsets with regulatory capacity, such as Th1 cells, are functionally impaired by SA. Our study will be an invention of novel anti-inflammatory agents to modulate the Th1/Th2 balance.
seeds on Th1/Th2 balance have been conducted to date. The purpose of this study is to determine whether SA may influence the direction of the Th1/Th2 balance as evaluated on the basis of splenic CD4+ T cell producing cytokine synthesis patterns in mice.
가설 설정
1. Effect of IFN-γ cytokine production in isolated CD4+ T cells.
2. Effect of IL-4 cytokine production in isolated CD4+ T cells.
To investigate the anti-inflammatory effect of SA, we examined the effect of SA on pro-inflammatory cytokines production. Effects of SA on peritoneal macrophages, there were marked reduction TNF-α cytokine production in high concentration (SA 100 μg/ml) compared to control group (Fig.
제안 방법
Real-time quantitative PCR was performed using the Applied Biosystems 7500 Fast Real-Time PCR system (Applied Biosystems, Foster, CA, US) with the following primer sequences: mouse transforming growth facto-β (TGF-β), 5'-TGGAGCAACATGTGGAACTC-3' and 5'-CTGCCGTACAACTCCAGTGA-3' β-actin, 5'-TGGAATCCTGTGGTCCATGAAAC- 3' and 5'-GTCACAGTCAGCTGTATAGGG-3'.
데이터처리
For statistical analysis of data, P-values were analyzed using a one-way analysis of variance (ANOVA) or unpaired Student’ t-test followed by Dunnett’ multiple comparison test (SPSS version 14.0 statistic software).
성능/효과
In conclusion, we demonstrated that SA inhibited Th1(IFN-γ) cytokine and proinflammatory cytokine(TNF-α) production in LPS-stimulated mouse peritoneal macrophages, and that these effects were mediated by TGFβ and Foxp3 gene dependently.
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