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CD25+ CD4+ T cells, expanded with dendritic cells presenting a single autoantigenic peptide, suppress autoimmune diabetes.

Tarbell KV, Yamazaki S, Olson K, Toy P, Steinman RM - J. Exp. Med. (2004)

Bottom Line: The expanded CD25+ CD4+ BDC2.5 T cells were effective even if administered 14 d after the diabetogenic T cells.Our data indicate that DCs can generate CD25+ CD4+ T cells that suppress autoimmune disease in vivo.This might be harnessed as a new avenue for immunotherapy, especially because CD25+ CD4+ regulatory cells responsive to a single autoantigen can inhibit diabetes mediated by reactivity to multiple antigens.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cellular Physiology and Immunology, The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA.

ABSTRACT
In the nonobese diabetic (NOD) mouse model of type 1 diabetes, the immune system recognizes many autoantigens expressed in pancreatic islet beta cells. To silence autoimmunity, we used dendritic cells (DCs) from NOD mice to expand CD25+ CD4+ suppressor T cells from BDC2.5 mice, which are specific for a single islet autoantigen. The expanded T cells were more suppressive in vitro than their freshly isolated counterparts, indicating that DCs from autoimmune mice can increase the number and function of antigen-specific, CD25+ CD4+ regulatory T cells. Importantly, only 5,000 expanded CD25+ CD4+ BDC2.5 T cells could block autoimmunity caused by diabetogenic T cells in NOD mice, whereas 10(5) polyclonal, CD25+ CD4+ T cells from NOD mice were inactive. When islets were examined in treated mice, insulitis development was blocked at early (3 wk) but not later (11 wk) time points. The expanded CD25+ CD4+ BDC2.5 T cells were effective even if administered 14 d after the diabetogenic T cells. Our data indicate that DCs can generate CD25+ CD4+ T cells that suppress autoimmune disease in vivo. This might be harnessed as a new avenue for immunotherapy, especially because CD25+ CD4+ regulatory cells responsive to a single autoantigen can inhibit diabetes mediated by reactivity to multiple antigens.

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DC-expanded CD25+ CD4+ T cells suppress proliferation better than unexpanded CD25+ CD4+ T cells. (A) CD25+ CD4+ T cells from NOD.BDC2.5 mice were expanded for 7 d with irradiated NOD DCs and BDC peptide and IL-2 as indicated. 104 freshly isolated, sorted CD25− CD4+ T cells from BDC2.5 mice were cultured with NOD spleen cells, 30 ng/ml BDC peptide, and either freshly sorted CD25+ CD4+ or the indicated DC-expanded CD25+ CD4+ populations, at the ratios indicated. After 72 h, proliferation was assessed by [3H]thymidine incorporation during a 12-h pulse. One representative result from at least three is shown. (B) Same as A, but both CD25+ and CD25− CD4+ T cells were isolated from NOD mice, and anti-CD3 was used as TCR stimulus instead of BDC peptide in both expansion and suppression cultures. One representative result from at least three is shown.
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fig4: DC-expanded CD25+ CD4+ T cells suppress proliferation better than unexpanded CD25+ CD4+ T cells. (A) CD25+ CD4+ T cells from NOD.BDC2.5 mice were expanded for 7 d with irradiated NOD DCs and BDC peptide and IL-2 as indicated. 104 freshly isolated, sorted CD25− CD4+ T cells from BDC2.5 mice were cultured with NOD spleen cells, 30 ng/ml BDC peptide, and either freshly sorted CD25+ CD4+ or the indicated DC-expanded CD25+ CD4+ populations, at the ratios indicated. After 72 h, proliferation was assessed by [3H]thymidine incorporation during a 12-h pulse. One representative result from at least three is shown. (B) Same as A, but both CD25+ and CD25− CD4+ T cells were isolated from NOD mice, and anti-CD3 was used as TCR stimulus instead of BDC peptide in both expansion and suppression cultures. One representative result from at least three is shown.

Mentions: To verify that the expanded CD25+ CD4+ T cells from BDC2.5 mice retained suppressive function, we used a standard in vitro suppression assay. We removed CD11c+ DCs from 7-d expansion cultures and added the T cells in different ratios to responder CD25− CD4+ T cells to measure the inhibition of CD25− CD4+ proliferation in response to BDC peptide presented by spleen APCs. Freshly isolated CD25+ CD4+ T cells, as well as CD25+ CD4+ T cells expanded with DCs and IL-2, were able to suppress, but only partially and at high doses, i.e., when mixed at a 1:2 ratio with CD25− CD4+ cells. In contrast, CD25+ CD4+ T cells expanded with BDC peptide (without or with IL-2) had stronger activity, showing suppression even at a ratio of 1 CD25+ CD4+ T cell for every 8 CD25− CD4+ cells (Fig. 4 A). We also tested the suppressive function of NOD CD25+ CD4+ T cells expanded with DCs and anti-CD3. Again, the T cells expanded with DCs and TCR stimulus suppressed proliferation by NOD CD25− CD4+ T cells approximately fourfold more efficiently than freshly isolated CD25+ CD4+ T cells (Fig. 4 B). Although freshly purified NOD CD25+ CD4+ T cells showed ∼75% suppression at a ratio of 8 responder cells for 1 CD25+ CD4+ T cell, NOD CD25+ CD4+ T cells expanded with DCs and anti-CD3 (with or without IL-2) showed similar suppression at a ratio of 32:1. Therefore, either polyclonal or monospecific CD25+ CD4+ T cells from NOD mice can be expanded with DCs and anti-CD3 or antigen, and they show approximately fourfold enhancement in suppressive function.


CD25+ CD4+ T cells, expanded with dendritic cells presenting a single autoantigenic peptide, suppress autoimmune diabetes.

Tarbell KV, Yamazaki S, Olson K, Toy P, Steinman RM - J. Exp. Med. (2004)

DC-expanded CD25+ CD4+ T cells suppress proliferation better than unexpanded CD25+ CD4+ T cells. (A) CD25+ CD4+ T cells from NOD.BDC2.5 mice were expanded for 7 d with irradiated NOD DCs and BDC peptide and IL-2 as indicated. 104 freshly isolated, sorted CD25− CD4+ T cells from BDC2.5 mice were cultured with NOD spleen cells, 30 ng/ml BDC peptide, and either freshly sorted CD25+ CD4+ or the indicated DC-expanded CD25+ CD4+ populations, at the ratios indicated. After 72 h, proliferation was assessed by [3H]thymidine incorporation during a 12-h pulse. One representative result from at least three is shown. (B) Same as A, but both CD25+ and CD25− CD4+ T cells were isolated from NOD mice, and anti-CD3 was used as TCR stimulus instead of BDC peptide in both expansion and suppression cultures. One representative result from at least three is shown.
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Related In: Results  -  Collection

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fig4: DC-expanded CD25+ CD4+ T cells suppress proliferation better than unexpanded CD25+ CD4+ T cells. (A) CD25+ CD4+ T cells from NOD.BDC2.5 mice were expanded for 7 d with irradiated NOD DCs and BDC peptide and IL-2 as indicated. 104 freshly isolated, sorted CD25− CD4+ T cells from BDC2.5 mice were cultured with NOD spleen cells, 30 ng/ml BDC peptide, and either freshly sorted CD25+ CD4+ or the indicated DC-expanded CD25+ CD4+ populations, at the ratios indicated. After 72 h, proliferation was assessed by [3H]thymidine incorporation during a 12-h pulse. One representative result from at least three is shown. (B) Same as A, but both CD25+ and CD25− CD4+ T cells were isolated from NOD mice, and anti-CD3 was used as TCR stimulus instead of BDC peptide in both expansion and suppression cultures. One representative result from at least three is shown.
Mentions: To verify that the expanded CD25+ CD4+ T cells from BDC2.5 mice retained suppressive function, we used a standard in vitro suppression assay. We removed CD11c+ DCs from 7-d expansion cultures and added the T cells in different ratios to responder CD25− CD4+ T cells to measure the inhibition of CD25− CD4+ proliferation in response to BDC peptide presented by spleen APCs. Freshly isolated CD25+ CD4+ T cells, as well as CD25+ CD4+ T cells expanded with DCs and IL-2, were able to suppress, but only partially and at high doses, i.e., when mixed at a 1:2 ratio with CD25− CD4+ cells. In contrast, CD25+ CD4+ T cells expanded with BDC peptide (without or with IL-2) had stronger activity, showing suppression even at a ratio of 1 CD25+ CD4+ T cell for every 8 CD25− CD4+ cells (Fig. 4 A). We also tested the suppressive function of NOD CD25+ CD4+ T cells expanded with DCs and anti-CD3. Again, the T cells expanded with DCs and TCR stimulus suppressed proliferation by NOD CD25− CD4+ T cells approximately fourfold more efficiently than freshly isolated CD25+ CD4+ T cells (Fig. 4 B). Although freshly purified NOD CD25+ CD4+ T cells showed ∼75% suppression at a ratio of 8 responder cells for 1 CD25+ CD4+ T cell, NOD CD25+ CD4+ T cells expanded with DCs and anti-CD3 (with or without IL-2) showed similar suppression at a ratio of 32:1. Therefore, either polyclonal or monospecific CD25+ CD4+ T cells from NOD mice can be expanded with DCs and anti-CD3 or antigen, and they show approximately fourfold enhancement in suppressive function.

Bottom Line: The expanded CD25+ CD4+ BDC2.5 T cells were effective even if administered 14 d after the diabetogenic T cells.Our data indicate that DCs can generate CD25+ CD4+ T cells that suppress autoimmune disease in vivo.This might be harnessed as a new avenue for immunotherapy, especially because CD25+ CD4+ regulatory cells responsive to a single autoantigen can inhibit diabetes mediated by reactivity to multiple antigens.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cellular Physiology and Immunology, The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA.

ABSTRACT
In the nonobese diabetic (NOD) mouse model of type 1 diabetes, the immune system recognizes many autoantigens expressed in pancreatic islet beta cells. To silence autoimmunity, we used dendritic cells (DCs) from NOD mice to expand CD25+ CD4+ suppressor T cells from BDC2.5 mice, which are specific for a single islet autoantigen. The expanded T cells were more suppressive in vitro than their freshly isolated counterparts, indicating that DCs from autoimmune mice can increase the number and function of antigen-specific, CD25+ CD4+ regulatory T cells. Importantly, only 5,000 expanded CD25+ CD4+ BDC2.5 T cells could block autoimmunity caused by diabetogenic T cells in NOD mice, whereas 10(5) polyclonal, CD25+ CD4+ T cells from NOD mice were inactive. When islets were examined in treated mice, insulitis development was blocked at early (3 wk) but not later (11 wk) time points. The expanded CD25+ CD4+ BDC2.5 T cells were effective even if administered 14 d after the diabetogenic T cells. Our data indicate that DCs can generate CD25+ CD4+ T cells that suppress autoimmune disease in vivo. This might be harnessed as a new avenue for immunotherapy, especially because CD25+ CD4+ regulatory cells responsive to a single autoantigen can inhibit diabetes mediated by reactivity to multiple antigens.

Show MeSH
Related in: MedlinePlus