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All-trans retinoic acid inhibits type 1 diabetes by T regulatory (Treg)-dependent suppression of interferon-gamma-producing T-cells without affecting Th17 cells.

Van YH, Lee WH, Ortiz S, Lee MH, Qin HJ, Liu CP - Diabetes (2008)

Bottom Line: Consistent with these results, ATRA reduced T-bet and STAT4 expression in T-cells and decreased islet-infiltrating CD8(+) T-cells, suppressing their activation and IFN-gamma/granzyme B expression.These results demonstrate that ATRA treatment promoted in vivo expansion of Treg cells and induced Treg cell-dependent immune tolerance by suppressing IFN-gamma-producing T-cells, without affecting Th17 cells.Our study also provides novel insights into how ATRA induces immune tolerance in vivo via its effects on Teff and Treg cells.

View Article: PubMed Central - PubMed

Affiliation: Division of Immunology, Beckman Research Institute, City of Hope, Duarte, California, USA.

ABSTRACT

Objective: All-trans retinoic acid (ATRA), a potent derivative of vitamin A, can regulate immune responses. However, its role in inducing immune tolerance associated with the prevention of islet inflammation and inhibition of type 1 diabetes remains unclear.

Research design and methods: We investigated the mechanisms underlying the potential immunoregulatory effect of ATRA on type 1 diabetes using an adoptive transfer animal model of the disease.

Results: Our data demonstrated that ATRA treatment inhibited diabetes in NOD mice with established insulitis. In addition, it suppressed interferon (IFN)-gamma-producing CD4(+) and CD8(+) T effector (Teff) cells and expanded T regulatory (Treg) cells in recipient mice transferred with diabetic NOD splenocytes, without affecting either interleukin (IL)-17--or IL-4-producing cells. Consistent with these results, ATRA reduced T-bet and STAT4 expression in T-cells and decreased islet-infiltrating CD8(+) T-cells, suppressing their activation and IFN-gamma/granzyme B expression. Depletion of CD4(+)CD25(+) Treg cells impaired the inhibitory effect of ATRA on islet-infiltrating T-cells and blocked its protective effect on diabetes. Therefore, ATRA treatment induced Treg cell-dependent immune tolerance by suppressing both CD4(+) and CD8(+) Teff cells while promoting Treg cell expansion.

Conclusions: These results demonstrate that ATRA treatment promoted in vivo expansion of Treg cells and induced Treg cell-dependent immune tolerance by suppressing IFN-gamma-producing T-cells, without affecting Th17 cells. Our study also provides novel insights into how ATRA induces immune tolerance in vivo via its effects on Teff and Treg cells.

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Related in: MedlinePlus

In vivo effects of ATRA treatment on IFN-γ and IL-17 production in T-cells of recipient mice. A: Intracellular staining of CD4+ T-cells with antibodies against IFN-γ, IL-17, or IL-4. Cells were isolated from pancreatic lymph nodes (PLN) or islets (ISL) of diabetic control or diabetes-free ATRA-treated recipient mice 4 weeks after cell transfer and surface-stained with anti-CD4 and then intracellularly stained with antibody for cytokines. Cells were electronically gated on CD4+ T-cells. B: Percentage of cells producing either IFN-γ (left) or IL-17 (right). n = 5. C: ATRA treatment suppresses in vivo production of IFN-γ by T-cells in response to anti-CD3 stimulation, detected by ELISA. Four weeks after adoptive transfer of diabetic NOD mouse splenocytes, diabetic control recipient mice and nondiabetic ATRA-treated recipient mice were intravenously injected with anti-CD3 (n = 3). D: ATRA treatment suppresses T-bet and STAT4 expression in T-cells. T-cells were isolated from the spleen of control or ATRA-treated recipient mice at 4 weeks post–cell transfer. Isolated T-cells were then stimulated with phorbol myristate acetate plus ionomycin. Western blot of cell lysates for T-bet and STAT4 expression (n = 3). (Please see http://dx.doi.org/10.2337/db08-1154 for a high-quality digital representation of this figure.)
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f5: In vivo effects of ATRA treatment on IFN-γ and IL-17 production in T-cells of recipient mice. A: Intracellular staining of CD4+ T-cells with antibodies against IFN-γ, IL-17, or IL-4. Cells were isolated from pancreatic lymph nodes (PLN) or islets (ISL) of diabetic control or diabetes-free ATRA-treated recipient mice 4 weeks after cell transfer and surface-stained with anti-CD4 and then intracellularly stained with antibody for cytokines. Cells were electronically gated on CD4+ T-cells. B: Percentage of cells producing either IFN-γ (left) or IL-17 (right). n = 5. C: ATRA treatment suppresses in vivo production of IFN-γ by T-cells in response to anti-CD3 stimulation, detected by ELISA. Four weeks after adoptive transfer of diabetic NOD mouse splenocytes, diabetic control recipient mice and nondiabetic ATRA-treated recipient mice were intravenously injected with anti-CD3 (n = 3). D: ATRA treatment suppresses T-bet and STAT4 expression in T-cells. T-cells were isolated from the spleen of control or ATRA-treated recipient mice at 4 weeks post–cell transfer. Isolated T-cells were then stimulated with phorbol myristate acetate plus ionomycin. Western blot of cell lysates for T-bet and STAT4 expression (n = 3). (Please see http://dx.doi.org/10.2337/db08-1154 for a high-quality digital representation of this figure.)

Mentions: We next investigated whether ATRA treatment had similar negative effects on Th1 and Th17 cells in vivo; in particular, we examined the percentage of IFN-γ–and IL-17–producing cells in recipient mice. Cells isolated from pancreatic lymph nodes and islets of recipient mice were analyzed by intracellular cytokine staining 4 weeks after cell transfer. The results showed that the percentage of IFN-γ–producing CD4+ cells was significantly reduced in both pancreatic lymph nodes and islets of ATRA-treated recipient mice compared with controls (Fig. 5A and B). However, unexpectedly, the percentage of IL-17–producing CD4+ cells was not significantly changed in these tissues, nor was there an effect on IL-4–producing CD4+ cells (Fig. 5A).


All-trans retinoic acid inhibits type 1 diabetes by T regulatory (Treg)-dependent suppression of interferon-gamma-producing T-cells without affecting Th17 cells.

Van YH, Lee WH, Ortiz S, Lee MH, Qin HJ, Liu CP - Diabetes (2008)

In vivo effects of ATRA treatment on IFN-γ and IL-17 production in T-cells of recipient mice. A: Intracellular staining of CD4+ T-cells with antibodies against IFN-γ, IL-17, or IL-4. Cells were isolated from pancreatic lymph nodes (PLN) or islets (ISL) of diabetic control or diabetes-free ATRA-treated recipient mice 4 weeks after cell transfer and surface-stained with anti-CD4 and then intracellularly stained with antibody for cytokines. Cells were electronically gated on CD4+ T-cells. B: Percentage of cells producing either IFN-γ (left) or IL-17 (right). n = 5. C: ATRA treatment suppresses in vivo production of IFN-γ by T-cells in response to anti-CD3 stimulation, detected by ELISA. Four weeks after adoptive transfer of diabetic NOD mouse splenocytes, diabetic control recipient mice and nondiabetic ATRA-treated recipient mice were intravenously injected with anti-CD3 (n = 3). D: ATRA treatment suppresses T-bet and STAT4 expression in T-cells. T-cells were isolated from the spleen of control or ATRA-treated recipient mice at 4 weeks post–cell transfer. Isolated T-cells were then stimulated with phorbol myristate acetate plus ionomycin. Western blot of cell lysates for T-bet and STAT4 expression (n = 3). (Please see http://dx.doi.org/10.2337/db08-1154 for a high-quality digital representation of this figure.)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2606864&req=5

f5: In vivo effects of ATRA treatment on IFN-γ and IL-17 production in T-cells of recipient mice. A: Intracellular staining of CD4+ T-cells with antibodies against IFN-γ, IL-17, or IL-4. Cells were isolated from pancreatic lymph nodes (PLN) or islets (ISL) of diabetic control or diabetes-free ATRA-treated recipient mice 4 weeks after cell transfer and surface-stained with anti-CD4 and then intracellularly stained with antibody for cytokines. Cells were electronically gated on CD4+ T-cells. B: Percentage of cells producing either IFN-γ (left) or IL-17 (right). n = 5. C: ATRA treatment suppresses in vivo production of IFN-γ by T-cells in response to anti-CD3 stimulation, detected by ELISA. Four weeks after adoptive transfer of diabetic NOD mouse splenocytes, diabetic control recipient mice and nondiabetic ATRA-treated recipient mice were intravenously injected with anti-CD3 (n = 3). D: ATRA treatment suppresses T-bet and STAT4 expression in T-cells. T-cells were isolated from the spleen of control or ATRA-treated recipient mice at 4 weeks post–cell transfer. Isolated T-cells were then stimulated with phorbol myristate acetate plus ionomycin. Western blot of cell lysates for T-bet and STAT4 expression (n = 3). (Please see http://dx.doi.org/10.2337/db08-1154 for a high-quality digital representation of this figure.)
Mentions: We next investigated whether ATRA treatment had similar negative effects on Th1 and Th17 cells in vivo; in particular, we examined the percentage of IFN-γ–and IL-17–producing cells in recipient mice. Cells isolated from pancreatic lymph nodes and islets of recipient mice were analyzed by intracellular cytokine staining 4 weeks after cell transfer. The results showed that the percentage of IFN-γ–producing CD4+ cells was significantly reduced in both pancreatic lymph nodes and islets of ATRA-treated recipient mice compared with controls (Fig. 5A and B). However, unexpectedly, the percentage of IL-17–producing CD4+ cells was not significantly changed in these tissues, nor was there an effect on IL-4–producing CD4+ cells (Fig. 5A).

Bottom Line: Consistent with these results, ATRA reduced T-bet and STAT4 expression in T-cells and decreased islet-infiltrating CD8(+) T-cells, suppressing their activation and IFN-gamma/granzyme B expression.These results demonstrate that ATRA treatment promoted in vivo expansion of Treg cells and induced Treg cell-dependent immune tolerance by suppressing IFN-gamma-producing T-cells, without affecting Th17 cells.Our study also provides novel insights into how ATRA induces immune tolerance in vivo via its effects on Teff and Treg cells.

View Article: PubMed Central - PubMed

Affiliation: Division of Immunology, Beckman Research Institute, City of Hope, Duarte, California, USA.

ABSTRACT

Objective: All-trans retinoic acid (ATRA), a potent derivative of vitamin A, can regulate immune responses. However, its role in inducing immune tolerance associated with the prevention of islet inflammation and inhibition of type 1 diabetes remains unclear.

Research design and methods: We investigated the mechanisms underlying the potential immunoregulatory effect of ATRA on type 1 diabetes using an adoptive transfer animal model of the disease.

Results: Our data demonstrated that ATRA treatment inhibited diabetes in NOD mice with established insulitis. In addition, it suppressed interferon (IFN)-gamma-producing CD4(+) and CD8(+) T effector (Teff) cells and expanded T regulatory (Treg) cells in recipient mice transferred with diabetic NOD splenocytes, without affecting either interleukin (IL)-17--or IL-4-producing cells. Consistent with these results, ATRA reduced T-bet and STAT4 expression in T-cells and decreased islet-infiltrating CD8(+) T-cells, suppressing their activation and IFN-gamma/granzyme B expression. Depletion of CD4(+)CD25(+) Treg cells impaired the inhibitory effect of ATRA on islet-infiltrating T-cells and blocked its protective effect on diabetes. Therefore, ATRA treatment induced Treg cell-dependent immune tolerance by suppressing both CD4(+) and CD8(+) Teff cells while promoting Treg cell expansion.

Conclusions: These results demonstrate that ATRA treatment promoted in vivo expansion of Treg cells and induced Treg cell-dependent immune tolerance by suppressing IFN-gamma-producing T-cells, without affecting Th17 cells. Our study also provides novel insights into how ATRA induces immune tolerance in vivo via its effects on Teff and Treg cells.

Show MeSH
Related in: MedlinePlus