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In vitro-expanded antigen-specific regulatory T cells suppress autoimmune diabetes.

Tang Q, Henriksen KJ, Bi M, Finger EB, Szot G, Ye J, Masteller EL, McDevitt H, Bonyhadi M, Bluestone JA - J. Exp. Med. (2004)

Bottom Line: Purified CD4+ CD25+ Tregs were expanded up to 200-fold in less than 2 wk in vitro using a combination of anti-CD3, anti-CD28, and interleukin 2.The expanded Tregs express a classical cell surface phenotype and function both in vitro and in vivo to suppress effector T cell functions.Most significantly, small numbers of antigen-specific Tregs can reverse diabetes after disease onset, suggesting a novel approach to cellular immunotherapy for autoimmunity.

View Article: PubMed Central - PubMed

Affiliation: UCSF Diabetes Center, University of California San Francisco, 94143, USA.

ABSTRACT
The low number of CD4+ CD25+ regulatory T cells (Tregs), their anergic phenotype, and diverse antigen specificity present major challenges to harnessing this potent tolerogenic population to treat autoimmunity and transplant rejection. In this study, we describe a robust method to expand antigen-specific Tregs from autoimmune-prone nonobese diabetic mice. Purified CD4+ CD25+ Tregs were expanded up to 200-fold in less than 2 wk in vitro using a combination of anti-CD3, anti-CD28, and interleukin 2. The expanded Tregs express a classical cell surface phenotype and function both in vitro and in vivo to suppress effector T cell functions. Most significantly, small numbers of antigen-specific Tregs can reverse diabetes after disease onset, suggesting a novel approach to cellular immunotherapy for autoimmunity.

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Reversal of diabetes with expanded Tregs. (A) NOD mice with chronic diabetes were transplanted with syngeneic islets under the kidney capsule. On the day of transplantation, some recipient mice received 5 × 106 NOD-expanded Tregs (n = 4) or 2 × 106 BDC2.5-expanded Tregs (n = 5), and the remaining mice (n = 3) were left untreated. Blood glucose level was monitored. All islet recipients normalized blood glucose within the first day after transplantation. Results are representative of two independent experiments. (B) NOD mice with new onset diabetes (blood glucose > 300 mg/dL, n = 7) were injected with 107 BDC2.5-expanded Tregs and blood glucose was monitored. Two consecutive readings of blood glucose of <250 mg/dL was considered remission of diabetes.
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fig7: Reversal of diabetes with expanded Tregs. (A) NOD mice with chronic diabetes were transplanted with syngeneic islets under the kidney capsule. On the day of transplantation, some recipient mice received 5 × 106 NOD-expanded Tregs (n = 4) or 2 × 106 BDC2.5-expanded Tregs (n = 5), and the remaining mice (n = 3) were left untreated. Blood glucose level was monitored. All islet recipients normalized blood glucose within the first day after transplantation. Results are representative of two independent experiments. (B) NOD mice with new onset diabetes (blood glucose > 300 mg/dL, n = 7) were injected with 107 BDC2.5-expanded Tregs and blood glucose was monitored. Two consecutive readings of blood glucose of <250 mg/dL was considered remission of diabetes.

Mentions: The ultimate utility of Treg therapy depends on an ability to treat individuals with ongoing disease. Thus, we examined the regulatory effects of expanded BDC2.5 Tregs in NOD mice that had been diabetic for at least 2 wk to ensure total endogenous islet cell destruction. Expanded BDC2.5 Tregs were transferred into diabetic NOD mice in conjunction with 500 syngeneic NOD islet transplant. Mice reverted to normoglycemia within 24–48 h after transplantation. However, unlike the control mice that rejected the transplanted islets within 2 wk, the transfer of 2 × 106 BDC2.5-expanded Tregs blocked rejection of the syngeneic islets, consistent with an ability of the suppressor cells to block ongoing autoimmunity in this setting. The transfer of 5 × 106 polyclonal NOD Tregs had no effect in this model. More significantly, the adoptive transfer of expanded BDC2.5 Tregs reversed diabetes in new-onset diabetic NOD mice (Fig. 7 B). Previous studies have shown that NOD mice diagnosed within the first week of hyperglycemia retain sufficient insulin-producing β cell activity, such that effective immunosuppression introduced at that time can reverse diabetes. To test the efficacy of expanded BDC2.5 Tregs in this setting, 107 Tregs were transferred into NOD mice diagnosed with disease based on recently elevated blood glucose levels (>300 mg/dL). The transferred Tregs reversed diabetes in 60% of the mice. Thus, the expanded Tregs were extremely effective in blocking and reversing diabetes in an ongoing autoimmune setting.


In vitro-expanded antigen-specific regulatory T cells suppress autoimmune diabetes.

Tang Q, Henriksen KJ, Bi M, Finger EB, Szot G, Ye J, Masteller EL, McDevitt H, Bonyhadi M, Bluestone JA - J. Exp. Med. (2004)

Reversal of diabetes with expanded Tregs. (A) NOD mice with chronic diabetes were transplanted with syngeneic islets under the kidney capsule. On the day of transplantation, some recipient mice received 5 × 106 NOD-expanded Tregs (n = 4) or 2 × 106 BDC2.5-expanded Tregs (n = 5), and the remaining mice (n = 3) were left untreated. Blood glucose level was monitored. All islet recipients normalized blood glucose within the first day after transplantation. Results are representative of two independent experiments. (B) NOD mice with new onset diabetes (blood glucose > 300 mg/dL, n = 7) were injected with 107 BDC2.5-expanded Tregs and blood glucose was monitored. Two consecutive readings of blood glucose of <250 mg/dL was considered remission of diabetes.
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Related In: Results  -  Collection

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fig7: Reversal of diabetes with expanded Tregs. (A) NOD mice with chronic diabetes were transplanted with syngeneic islets under the kidney capsule. On the day of transplantation, some recipient mice received 5 × 106 NOD-expanded Tregs (n = 4) or 2 × 106 BDC2.5-expanded Tregs (n = 5), and the remaining mice (n = 3) were left untreated. Blood glucose level was monitored. All islet recipients normalized blood glucose within the first day after transplantation. Results are representative of two independent experiments. (B) NOD mice with new onset diabetes (blood glucose > 300 mg/dL, n = 7) were injected with 107 BDC2.5-expanded Tregs and blood glucose was monitored. Two consecutive readings of blood glucose of <250 mg/dL was considered remission of diabetes.
Mentions: The ultimate utility of Treg therapy depends on an ability to treat individuals with ongoing disease. Thus, we examined the regulatory effects of expanded BDC2.5 Tregs in NOD mice that had been diabetic for at least 2 wk to ensure total endogenous islet cell destruction. Expanded BDC2.5 Tregs were transferred into diabetic NOD mice in conjunction with 500 syngeneic NOD islet transplant. Mice reverted to normoglycemia within 24–48 h after transplantation. However, unlike the control mice that rejected the transplanted islets within 2 wk, the transfer of 2 × 106 BDC2.5-expanded Tregs blocked rejection of the syngeneic islets, consistent with an ability of the suppressor cells to block ongoing autoimmunity in this setting. The transfer of 5 × 106 polyclonal NOD Tregs had no effect in this model. More significantly, the adoptive transfer of expanded BDC2.5 Tregs reversed diabetes in new-onset diabetic NOD mice (Fig. 7 B). Previous studies have shown that NOD mice diagnosed within the first week of hyperglycemia retain sufficient insulin-producing β cell activity, such that effective immunosuppression introduced at that time can reverse diabetes. To test the efficacy of expanded BDC2.5 Tregs in this setting, 107 Tregs were transferred into NOD mice diagnosed with disease based on recently elevated blood glucose levels (>300 mg/dL). The transferred Tregs reversed diabetes in 60% of the mice. Thus, the expanded Tregs were extremely effective in blocking and reversing diabetes in an ongoing autoimmune setting.

Bottom Line: Purified CD4+ CD25+ Tregs were expanded up to 200-fold in less than 2 wk in vitro using a combination of anti-CD3, anti-CD28, and interleukin 2.The expanded Tregs express a classical cell surface phenotype and function both in vitro and in vivo to suppress effector T cell functions.Most significantly, small numbers of antigen-specific Tregs can reverse diabetes after disease onset, suggesting a novel approach to cellular immunotherapy for autoimmunity.

View Article: PubMed Central - PubMed

Affiliation: UCSF Diabetes Center, University of California San Francisco, 94143, USA.

ABSTRACT
The low number of CD4+ CD25+ regulatory T cells (Tregs), their anergic phenotype, and diverse antigen specificity present major challenges to harnessing this potent tolerogenic population to treat autoimmunity and transplant rejection. In this study, we describe a robust method to expand antigen-specific Tregs from autoimmune-prone nonobese diabetic mice. Purified CD4+ CD25+ Tregs were expanded up to 200-fold in less than 2 wk in vitro using a combination of anti-CD3, anti-CD28, and interleukin 2. The expanded Tregs express a classical cell surface phenotype and function both in vitro and in vivo to suppress effector T cell functions. Most significantly, small numbers of antigen-specific Tregs can reverse diabetes after disease onset, suggesting a novel approach to cellular immunotherapy for autoimmunity.

Show MeSH
Related in: MedlinePlus