Polyunsaturated fatty acids promote the expansion of myeloid-derived suppressor cells by activating the JAK/STAT3 pathway
Article first published online: 29 AUG 2013
© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
European Journal of Immunology
Volume 43, Issue 11, pages 2943–2955, November 2013
How to Cite
Yan, D., Yang, Q., Shi, M., Zhong, L., Wu, C., Meng, T., Yin, H. and Zhou, J. (2013), Polyunsaturated fatty acids promote the expansion of myeloid-derived suppressor cells by activating the JAK/STAT3 pathway. Eur. J. Immunol., 43: 2943–2955. doi: 10.1002/eji.201343472
- Issue published online: 20 NOV 2013
- Article first published online: 29 AUG 2013
- Accepted manuscript online: 30 JUL 2013 02:17AM EST
- Manuscript Accepted: 26 JUL 2013
- Manuscript Revised: 25 JUN 2013
- Manuscript Received: 21 FEB 2013
- National Key Basic Research Program of China. Grant Number: 2012CB524900
- National Natural Science Foundation of China. Grant Numbers: 81072397, 31270921
- Guangdong Innovative Research Team Program. Grant Number: 2009010058
- Natural Science Foundation of Guangdong. Grant Number: S2011020006072
As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re-organized for online delivery, but are not copy-edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.
Figure 1: Mouse BM cells were cultured in medium containing GM-CSF (20 ng/mL) and IL-6 (20 ng/mL) for 6 days in the presence of 100 μM ALA or LA,ethanol was used as control. The proportion of MDSCs (Gr-1+CD11b+) was evaluated by flow cytometry. Left panels are from a single experiment, the right graph represents mean ± SEM of 6 samples pooled from 3 independent experiments. **p < 0.01: statistically different from control, unpaired t test was used.
Figure 2: BM cells from C57BL/6 mice were cultured in GM-CSF (20 ng/mL) for 6 days in the presence of 100 μM ALA or LA; ethanol was used as control.CD3+ T cells from BALB/c mice were stimulated with ConA, then cocultured with MDSCs isolated from the above cultured BM at different ratios for 3 days. T-cell proliferation was evaluated by CFSE dilution. Unstimulated T cells were used as a negative control. (A) The suppressive function of LA-induced MDSCs and control MDSCs. Left panels are from a single experiment; the right panels represent mean ± SEM of 6 samples pooled from 3 independent experiments. *p < 0.05, **p < 0.01: statistically significance compared with ConA-stimulated T cells.(B) The comparison in suppressive activity between PUFA-induced MDSCs and control MDSCs. Data represent mean ± SEM of 6 samples pooled from 3 independent experiments. *p < 0.05, **p < 0.01: compared with control MDSCs, unpaired t test was used.
Figure 3: Effects of different L-arginine related inhibitors on T-cell proliferation. CD3+ T cells from mouse spleen were stimulated with ConA for 3 days with treatments as indicated. T-cell proliferation was evaluated by CFSE dilution. Unstimulated T cells were used as a negative control. nor-NOHA(100 μM): arginase inhibitor; L-arginine(1 mM); L-NMMA (100 μM): iNOS inhibitor; NAC (1 mM): ROS inhibitor. Means ± SEM of 6 samples from 3 independent experiments are shown, unpaired t test was used.
Figure 4: Mice were fed diets enriched with ALA (LOD, linseed oil diet) or control diet (RD, regular diet) for 3 months. Gr1+CD11b+ cells were purified by flow cytometric sorting from BM and spleen, and the amounts of p-STAT3, STAT3, p47phox, S100A8/A9 proteins were determined by western blotting. Data are representative of 3 independent experiments.
Figure 5: Effect of PUFA on COX2 expression and the impact of COX2 inhibitor on PUFA-MDSC function. Mouse BM cells were cultured in GM-CSF (20 ng/mL) for 6 days with different treatments as indicated. NAC: ROS inhibitor, Nimesulide (Nim):COX2 inhibitor, ethanol was used as control. Gr1+CD11b+ cells were isolated from the cultured BM cells and subjected to the following experiments. (A) Expression of COX1 and COX2 was determined by qRT-PCR. (B) ROS level was determined by CM-H2DCFDA labeling.(C) Allogeneic MLR. MDSCs from the above cultures were cocultured with allogeneic CD3+ T cells stimulated with ConA, at 2:1 ratio for 3 days. T-cell proliferation was evaluated by CFSE dilution. Unstimulated T cells were used as a negative control. (A-C) Means ± SEM from 3 independent experiments are shown.
Table 1:Sequences of primers used in this study.
Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.