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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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Related in: MedlinePlus

Prevention of autoimmune diabetes in NOD.CD28−/− mice with BDC2.5-expanded Tregs. 5-wk-old prediabetic NOD.CD28−/− mice were injected with 5 × 105 BDC2.5-expanded Tregs (n = 3) or left untreated (n = 4). The development of diabetes was monitored and blood glucose levels of individual mice were plotted. Results are representative of at least five independent experiments.
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fig6: Prevention of autoimmune diabetes in NOD.CD28−/− mice with BDC2.5-expanded Tregs. 5-wk-old prediabetic NOD.CD28−/− mice were injected with 5 × 105 BDC2.5-expanded Tregs (n = 3) or left untreated (n = 4). The development of diabetes was monitored and blood glucose levels of individual mice were plotted. Results are representative of at least five independent experiments.

Mentions: Although there are multiple models demonstrating the immunoregulatory activity of Tregs, many of the systems are based on adoptive transfer models that take advantage of lymphopenic mice to enhance Treg proliferation (8, 21–24). Questions have been raised whether disease suppression observed in lymphopenic settings after Treg transfer is due to active regulation or a side efffect of competition for “space” (37, 38). Therefore, we examined the ability of the expanded Tregs to prevent diabetes in a nonlymphopenic animal model. Previous studies have shown that CD28−/− NOD mice have normal numbers of T cells and Th1 responses. In fact, these mice develop exacerbated autoimmunity due to a deficiency in Th2 and Tregs, which were shown to be exquisitely CD28 dependent (8, 21–24). Thus, we examined whether wild-type expanded BDC2.5 Tregs transferred into CD28−/− NOD mice could delay or prevent onset of disease. 5 × 105 Tregs were transferred into 5-wk-old CD28−/− NOD mice and monitored for diabetes (Fig. 6). The transfer of expanded BDC2.5 Tregs prevented the development of diabetes in 100% of mice followed for as long as 20 wk after transfer. In contrast, the transfer of similar numbers of expanded NOD Tregs had no effect on disease incidence (unpublished data). These results suggest that the antigen-specific expanded Tregs functioned in vivo in the face of a fully functional pathogenic T cell response.


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)

Prevention of autoimmune diabetes in NOD.CD28−/− mice with BDC2.5-expanded Tregs. 5-wk-old prediabetic NOD.CD28−/− mice were injected with 5 × 105 BDC2.5-expanded Tregs (n = 3) or left untreated (n = 4). The development of diabetes was monitored and blood glucose levels of individual mice were plotted. Results are representative of at least five independent experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2211775&req=5

fig6: Prevention of autoimmune diabetes in NOD.CD28−/− mice with BDC2.5-expanded Tregs. 5-wk-old prediabetic NOD.CD28−/− mice were injected with 5 × 105 BDC2.5-expanded Tregs (n = 3) or left untreated (n = 4). The development of diabetes was monitored and blood glucose levels of individual mice were plotted. Results are representative of at least five independent experiments.
Mentions: Although there are multiple models demonstrating the immunoregulatory activity of Tregs, many of the systems are based on adoptive transfer models that take advantage of lymphopenic mice to enhance Treg proliferation (8, 21–24). Questions have been raised whether disease suppression observed in lymphopenic settings after Treg transfer is due to active regulation or a side efffect of competition for “space” (37, 38). Therefore, we examined the ability of the expanded Tregs to prevent diabetes in a nonlymphopenic animal model. Previous studies have shown that CD28−/− NOD mice have normal numbers of T cells and Th1 responses. In fact, these mice develop exacerbated autoimmunity due to a deficiency in Th2 and Tregs, which were shown to be exquisitely CD28 dependent (8, 21–24). Thus, we examined whether wild-type expanded BDC2.5 Tregs transferred into CD28−/− NOD mice could delay or prevent onset of disease. 5 × 105 Tregs were transferred into 5-wk-old CD28−/− NOD mice and monitored for diabetes (Fig. 6). The transfer of expanded BDC2.5 Tregs prevented the development of diabetes in 100% of mice followed for as long as 20 wk after transfer. In contrast, the transfer of similar numbers of expanded NOD Tregs had no effect on disease incidence (unpublished data). These results suggest that the antigen-specific expanded Tregs functioned in vivo in the face of a fully functional pathogenic T cell response.

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