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Vitamin A Impairs the Reprogramming of Tregs into IL-17-Producing Cells during Intestinal Inflammation.

Tejón G, Manríquez V, De Calisto J, Flores-Santibáñez F, Hidalgo Y, Crisóstomo N, Fernández D, Sauma D, Mora JR, Bono MR, Rosemblatt M - Biomed Res Int (2015)

Bottom Line: Our results show that intestinal inflammation induces the reprogramming of iTreg cells into IL-17-producing cells and that vitamin A restrains reprogramming in the gut.We also demonstrate that the presence of IL-2 during the in vitro generation of iTreg cells confers resistance to Th17 conversion but that IL-2 and retinoic acid (RA) cooperate to maintain Foxp3 expression following stimulation under Th17-polarizing conditions.Additionally, although IL-2 and RA differentially regulate the expression of different Treg cell suppressive markers, Treg cells generated under different polarizing conditions present similar suppressive capacity.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Biología, Facultad de Ciencias, Universidad de Chile, 7800024 Santiago, Chile.

ABSTRACT
Maintaining the identity of Foxp3(+) regulatory T cells (Tregs) is critical for controlling immune responses in the gut, where an imbalance between Tregs and T effector cells has been linked to inflammatory bowel disease. Accumulating evidence suggests that Tregs can convert into Th17 cells and acquire an inflammatory phenotype. In this study, we used an adoptive transfer model of Ag-specific T cells to study the contribution of different factors to the reprogramming of in vitro-generated Treg cells (iTreg) into IL-17-producing cells in a mouse model of gut inflammation in vivo. Our results show that intestinal inflammation induces the reprogramming of iTreg cells into IL-17-producing cells and that vitamin A restrains reprogramming in the gut. We also demonstrate that the presence of IL-2 during the in vitro generation of iTreg cells confers resistance to Th17 conversion but that IL-2 and retinoic acid (RA) cooperate to maintain Foxp3 expression following stimulation under Th17-polarizing conditions. Additionally, although IL-2 and RA differentially regulate the expression of different Treg cell suppressive markers, Treg cells generated under different polarizing conditions present similar suppressive capacity.

No MeSH data available.


Related in: MedlinePlus

IL-2 confers iTreg resistance to Th17 conversion, while RA and IL-2 cooperate to sustain Foxp3 expression following stimulation under Th17-polarizing conditions. FACS-sorted antigen-specific iTreg cells generated in the presence of TGF-β plus IL-2 (TIL-2 Treg), plus RA (TRA Treg), or plus both RA and IL-2 (TILRA Treg) were recultured under Th17-polarizing conditions with IL-6, TGF-β1, IL-1β, and α-IFNγ. Six days later, the cells were collected for intracellular IL-17 and Foxp3-GFP staining. (a) Representative staining showing the percentage of Foxp3-GFP expression versus IL-17 content. (b) Values indicate means ± SEM of four separate experiments. Repeated measure ANOVA with Bonferroni's post test ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.
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fig3: IL-2 confers iTreg resistance to Th17 conversion, while RA and IL-2 cooperate to sustain Foxp3 expression following stimulation under Th17-polarizing conditions. FACS-sorted antigen-specific iTreg cells generated in the presence of TGF-β plus IL-2 (TIL-2 Treg), plus RA (TRA Treg), or plus both RA and IL-2 (TILRA Treg) were recultured under Th17-polarizing conditions with IL-6, TGF-β1, IL-1β, and α-IFNγ. Six days later, the cells were collected for intracellular IL-17 and Foxp3-GFP staining. (a) Representative staining showing the percentage of Foxp3-GFP expression versus IL-17 content. (b) Values indicate means ± SEM of four separate experiments. Repeated measure ANOVA with Bonferroni's post test ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.

Mentions: Based on reports that established that Foxp3 expression is a key factor for the generation and function of Treg cells [31], we decided to investigate whether RA and IL-2 affect the stability of Foxp3. To address this aim, iTreg cells generated under the different conditions were sorted and restimulated for 6 days under Th17-polarizing conditions. The cells were then analyzed by flow cytometry to assess the expression of Foxp-3 and IL-17 production. We observed that when Treg cells that were generated in the presence of both IL-2 and RA (TILRA) were placed under Th17-polarizing conditions, a higher number of cells retained Foxp3+ expression (42.0 ± 10.9%) compared with Tregs generated in the presence of IL-2 alone (22.5 ± 9.0%) or RA alone (26.0 ± 9.1%) (Figure 3). Furthermore, the results show that, in the absence of IL-2 (TRA Treg), Treg cells convert to a Th17 phenotype (56.1 ± 18.0%) at a higher frequency compared with TILRA (31.0 ± 16.0%) or TIL-2 (48.0 ± 18.0%) Treg cells. Taken together, these data suggest that the simultaneous presence of RA and IL-2 contributed to a more stable Foxp3 expression, which may favor Treg stability under inflammatory conditions.


Vitamin A Impairs the Reprogramming of Tregs into IL-17-Producing Cells during Intestinal Inflammation.

Tejón G, Manríquez V, De Calisto J, Flores-Santibáñez F, Hidalgo Y, Crisóstomo N, Fernández D, Sauma D, Mora JR, Bono MR, Rosemblatt M - Biomed Res Int (2015)

IL-2 confers iTreg resistance to Th17 conversion, while RA and IL-2 cooperate to sustain Foxp3 expression following stimulation under Th17-polarizing conditions. FACS-sorted antigen-specific iTreg cells generated in the presence of TGF-β plus IL-2 (TIL-2 Treg), plus RA (TRA Treg), or plus both RA and IL-2 (TILRA Treg) were recultured under Th17-polarizing conditions with IL-6, TGF-β1, IL-1β, and α-IFNγ. Six days later, the cells were collected for intracellular IL-17 and Foxp3-GFP staining. (a) Representative staining showing the percentage of Foxp3-GFP expression versus IL-17 content. (b) Values indicate means ± SEM of four separate experiments. Repeated measure ANOVA with Bonferroni's post test ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4637025&req=5

fig3: IL-2 confers iTreg resistance to Th17 conversion, while RA and IL-2 cooperate to sustain Foxp3 expression following stimulation under Th17-polarizing conditions. FACS-sorted antigen-specific iTreg cells generated in the presence of TGF-β plus IL-2 (TIL-2 Treg), plus RA (TRA Treg), or plus both RA and IL-2 (TILRA Treg) were recultured under Th17-polarizing conditions with IL-6, TGF-β1, IL-1β, and α-IFNγ. Six days later, the cells were collected for intracellular IL-17 and Foxp3-GFP staining. (a) Representative staining showing the percentage of Foxp3-GFP expression versus IL-17 content. (b) Values indicate means ± SEM of four separate experiments. Repeated measure ANOVA with Bonferroni's post test ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.
Mentions: Based on reports that established that Foxp3 expression is a key factor for the generation and function of Treg cells [31], we decided to investigate whether RA and IL-2 affect the stability of Foxp3. To address this aim, iTreg cells generated under the different conditions were sorted and restimulated for 6 days under Th17-polarizing conditions. The cells were then analyzed by flow cytometry to assess the expression of Foxp-3 and IL-17 production. We observed that when Treg cells that were generated in the presence of both IL-2 and RA (TILRA) were placed under Th17-polarizing conditions, a higher number of cells retained Foxp3+ expression (42.0 ± 10.9%) compared with Tregs generated in the presence of IL-2 alone (22.5 ± 9.0%) or RA alone (26.0 ± 9.1%) (Figure 3). Furthermore, the results show that, in the absence of IL-2 (TRA Treg), Treg cells convert to a Th17 phenotype (56.1 ± 18.0%) at a higher frequency compared with TILRA (31.0 ± 16.0%) or TIL-2 (48.0 ± 18.0%) Treg cells. Taken together, these data suggest that the simultaneous presence of RA and IL-2 contributed to a more stable Foxp3 expression, which may favor Treg stability under inflammatory conditions.

Bottom Line: Our results show that intestinal inflammation induces the reprogramming of iTreg cells into IL-17-producing cells and that vitamin A restrains reprogramming in the gut.We also demonstrate that the presence of IL-2 during the in vitro generation of iTreg cells confers resistance to Th17 conversion but that IL-2 and retinoic acid (RA) cooperate to maintain Foxp3 expression following stimulation under Th17-polarizing conditions.Additionally, although IL-2 and RA differentially regulate the expression of different Treg cell suppressive markers, Treg cells generated under different polarizing conditions present similar suppressive capacity.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Biología, Facultad de Ciencias, Universidad de Chile, 7800024 Santiago, Chile.

ABSTRACT
Maintaining the identity of Foxp3(+) regulatory T cells (Tregs) is critical for controlling immune responses in the gut, where an imbalance between Tregs and T effector cells has been linked to inflammatory bowel disease. Accumulating evidence suggests that Tregs can convert into Th17 cells and acquire an inflammatory phenotype. In this study, we used an adoptive transfer model of Ag-specific T cells to study the contribution of different factors to the reprogramming of in vitro-generated Treg cells (iTreg) into IL-17-producing cells in a mouse model of gut inflammation in vivo. Our results show that intestinal inflammation induces the reprogramming of iTreg cells into IL-17-producing cells and that vitamin A restrains reprogramming in the gut. We also demonstrate that the presence of IL-2 during the in vitro generation of iTreg cells confers resistance to Th17 conversion but that IL-2 and retinoic acid (RA) cooperate to maintain Foxp3 expression following stimulation under Th17-polarizing conditions. Additionally, although IL-2 and RA differentially regulate the expression of different Treg cell suppressive markers, Treg cells generated under different polarizing conditions present similar suppressive capacity.

No MeSH data available.


Related in: MedlinePlus