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Mesenchymal stem cells-regulated Treg cells suppress colitis-associated colorectal cancer.

Tang RJ, Shen SN, Zhao XY, Nie YZ, Xu YJ, Ren J, Lv MM, Hou YY, Wang TT - Stem Cell Res Ther (2015)

Bottom Line: Colitis-associated colorectal cancer model was induced using azoxymethane (AOM) and dextran sulfate sodium (DSS) and MSCs were injected intravenously twice.Naïve T cells and Jurkat cells were co-cultured with MSCs and the effect of MSCs on Treg cells differentiation was evaluated.These results proved that MSCs could migrate to colon tissues and induce the differentiation of Treg cells via Smad2 as so to inhibit the colitis and suppress the development of CAC.

View Article: PubMed Central - PubMed

Affiliation: The State Key Laboratory of Pharmaceutical Biotechnology, Division of Immunology, Medical School, Nanjing University, Nanjing, 210093, China. trjtrjtrj@163.com.

ABSTRACT

Introduction: Previous studies have produced controversial results regarding whether mesenchymal stem cells (MSCs) promote or inhibit tumor development. Given the dual role of MSCs in inflammation and cancer, in this study the colitis-associated colorectal cancer (CAC) model was used to examine whether umbilical cord tissue-derived MSCs could prevent neoplasm by inhibiting chronic inflammation.

Methods: MSCs were obtained and identified using flow cytometry. Colitis-associated colorectal cancer model was induced using azoxymethane (AOM) and dextran sulfate sodium (DSS) and MSCs were injected intravenously twice. Levels of immune cells in mesenteric lymph node including regulatory T (Treg) cells were detected using flow cytometry. Naïve T cells and Jurkat cells were co-cultured with MSCs and the effect of MSCs on Treg cells differentiation was evaluated.

Results: After injection through tail vein, MSCs could migrate to colon and suppress colitis-related neoplasm. This tumor suppressive effect was characterized by longer colon length, decreased tumor numbers and decreased expression of Ki-67. Moreover, MSCs alleviated the pathology of inflammation in the colitis stage of CAC model and inhibited inflammation cytokines both in colon and serum. Furthermore, Treg cells were accumulated in mesenteric lymph node of MSCs-treated mice while the percentage of T helper cells 2 (Th2) and Th17 were not changed. Of note, MSCs secreted transforming growth factor-β (TGF-β) enhanced the induction of Treg cells from naïve T cells. The conditioned medium of MSCs also activated Smad2 signaling, which has been reported to regulate Treg cells.

Conclusions: These results proved that MSCs could migrate to colon tissues and induce the differentiation of Treg cells via Smad2 as so to inhibit the colitis and suppress the development of CAC.

No MeSH data available.


Related in: MedlinePlus

The effect of MSCs on the accumulation of CD4+CD25+FoxP3+Treg cells. (A) Flow cytometry analysis of the mean percentage of CD4+T cells and CD8+T cells in mesenteric lymph. (B) Flow cytometry analyzed the mean percentage of Th2, Th17 and Treg cells in mesenteric lymph. Values are expressed as means ± SEM. *P <0.05. Colons collected at day 33 (C) and day 70 (D) stained with Foxp3 examined by immunohistochemistry. MSCs, mesenchymal stem cells; SEM, standard error of the mean; Treg cells, regulatory T cells.
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Fig5: The effect of MSCs on the accumulation of CD4+CD25+FoxP3+Treg cells. (A) Flow cytometry analysis of the mean percentage of CD4+T cells and CD8+T cells in mesenteric lymph. (B) Flow cytometry analyzed the mean percentage of Th2, Th17 and Treg cells in mesenteric lymph. Values are expressed as means ± SEM. *P <0.05. Colons collected at day 33 (C) and day 70 (D) stained with Foxp3 examined by immunohistochemistry. MSCs, mesenchymal stem cells; SEM, standard error of the mean; Treg cells, regulatory T cells.

Mentions: To detect the regulatory effect of MSCs on immune cells, flow cytometry was used to analyze changes of adaptive immune cells in mesenteric lymph nodes after treatment with MSCs for 33 days (colitis stage). As shown in Figure 5A, the mean percentage of CD4+T cells and CD8+T cells reflected no difference among groups as well as Th2 and Th17 cells. As is well known, forkhead box P3 (FoxP3) is the transcription factor of Treg cells. Notably, CD4+CD25+FoxP3+ Treg cells were significantly up-regulated by MSC treatment (Figure 5B, P < 0.05). These phenomena suggest that MSC may just induce the accumulation of Treg cells in mesenteric lymph nodes to suppress excessive inflammation after MSCs migrate to the colon. To further confirm the involvement of Treg cells in intestinal inflammation, FoxP3 stained by immunohistochemistry was used to analyze whether Treg cells infiltrated to the stroma and epithelia of the intestine. Interestingly, according to the density of FoxP3, we found that Treg cells infiltrated in intestinal stroma more in the MSC treated group than in the untreated group (Figure 5C). We also determined the density of Treg cells at day 70 (tumor stage). In contrast, the results showed that the infiltrated Treg cells were found less in the MSCs group compared to the neoplasm in the untreated group (Figure 5D). Therefore, these results indicated that MSCs indeed were involved in inducing Treg cells to suppress colitis.Figure 5


Mesenchymal stem cells-regulated Treg cells suppress colitis-associated colorectal cancer.

Tang RJ, Shen SN, Zhao XY, Nie YZ, Xu YJ, Ren J, Lv MM, Hou YY, Wang TT - Stem Cell Res Ther (2015)

The effect of MSCs on the accumulation of CD4+CD25+FoxP3+Treg cells. (A) Flow cytometry analysis of the mean percentage of CD4+T cells and CD8+T cells in mesenteric lymph. (B) Flow cytometry analyzed the mean percentage of Th2, Th17 and Treg cells in mesenteric lymph. Values are expressed as means ± SEM. *P <0.05. Colons collected at day 33 (C) and day 70 (D) stained with Foxp3 examined by immunohistochemistry. MSCs, mesenchymal stem cells; SEM, standard error of the mean; Treg cells, regulatory T cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4414289&req=5

Fig5: The effect of MSCs on the accumulation of CD4+CD25+FoxP3+Treg cells. (A) Flow cytometry analysis of the mean percentage of CD4+T cells and CD8+T cells in mesenteric lymph. (B) Flow cytometry analyzed the mean percentage of Th2, Th17 and Treg cells in mesenteric lymph. Values are expressed as means ± SEM. *P <0.05. Colons collected at day 33 (C) and day 70 (D) stained with Foxp3 examined by immunohistochemistry. MSCs, mesenchymal stem cells; SEM, standard error of the mean; Treg cells, regulatory T cells.
Mentions: To detect the regulatory effect of MSCs on immune cells, flow cytometry was used to analyze changes of adaptive immune cells in mesenteric lymph nodes after treatment with MSCs for 33 days (colitis stage). As shown in Figure 5A, the mean percentage of CD4+T cells and CD8+T cells reflected no difference among groups as well as Th2 and Th17 cells. As is well known, forkhead box P3 (FoxP3) is the transcription factor of Treg cells. Notably, CD4+CD25+FoxP3+ Treg cells were significantly up-regulated by MSC treatment (Figure 5B, P < 0.05). These phenomena suggest that MSC may just induce the accumulation of Treg cells in mesenteric lymph nodes to suppress excessive inflammation after MSCs migrate to the colon. To further confirm the involvement of Treg cells in intestinal inflammation, FoxP3 stained by immunohistochemistry was used to analyze whether Treg cells infiltrated to the stroma and epithelia of the intestine. Interestingly, according to the density of FoxP3, we found that Treg cells infiltrated in intestinal stroma more in the MSC treated group than in the untreated group (Figure 5C). We also determined the density of Treg cells at day 70 (tumor stage). In contrast, the results showed that the infiltrated Treg cells were found less in the MSCs group compared to the neoplasm in the untreated group (Figure 5D). Therefore, these results indicated that MSCs indeed were involved in inducing Treg cells to suppress colitis.Figure 5

Bottom Line: Colitis-associated colorectal cancer model was induced using azoxymethane (AOM) and dextran sulfate sodium (DSS) and MSCs were injected intravenously twice.Naïve T cells and Jurkat cells were co-cultured with MSCs and the effect of MSCs on Treg cells differentiation was evaluated.These results proved that MSCs could migrate to colon tissues and induce the differentiation of Treg cells via Smad2 as so to inhibit the colitis and suppress the development of CAC.

View Article: PubMed Central - PubMed

Affiliation: The State Key Laboratory of Pharmaceutical Biotechnology, Division of Immunology, Medical School, Nanjing University, Nanjing, 210093, China. trjtrjtrj@163.com.

ABSTRACT

Introduction: Previous studies have produced controversial results regarding whether mesenchymal stem cells (MSCs) promote or inhibit tumor development. Given the dual role of MSCs in inflammation and cancer, in this study the colitis-associated colorectal cancer (CAC) model was used to examine whether umbilical cord tissue-derived MSCs could prevent neoplasm by inhibiting chronic inflammation.

Methods: MSCs were obtained and identified using flow cytometry. Colitis-associated colorectal cancer model was induced using azoxymethane (AOM) and dextran sulfate sodium (DSS) and MSCs were injected intravenously twice. Levels of immune cells in mesenteric lymph node including regulatory T (Treg) cells were detected using flow cytometry. Naïve T cells and Jurkat cells were co-cultured with MSCs and the effect of MSCs on Treg cells differentiation was evaluated.

Results: After injection through tail vein, MSCs could migrate to colon and suppress colitis-related neoplasm. This tumor suppressive effect was characterized by longer colon length, decreased tumor numbers and decreased expression of Ki-67. Moreover, MSCs alleviated the pathology of inflammation in the colitis stage of CAC model and inhibited inflammation cytokines both in colon and serum. Furthermore, Treg cells were accumulated in mesenteric lymph node of MSCs-treated mice while the percentage of T helper cells 2 (Th2) and Th17 were not changed. Of note, MSCs secreted transforming growth factor-β (TGF-β) enhanced the induction of Treg cells from naïve T cells. The conditioned medium of MSCs also activated Smad2 signaling, which has been reported to regulate Treg cells.

Conclusions: These results proved that MSCs could migrate to colon tissues and induce the differentiation of Treg cells via Smad2 as so to inhibit the colitis and suppress the development of CAC.

No MeSH data available.


Related in: MedlinePlus