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DCIR2 + cDC2 DCs and Zbtb32 Restore CD4 + T-Cell Tolerance and Inhibit Diabetes

View Article: PubMed Central - PubMed

ABSTRACT

During autoimmunity, the normal ability of dendritic cells (DCs) to induce T-cell tolerance is disrupted; therefore, autoimmune disease therapies based on cell types and molecular pathways that elicit tolerance in the steady state may not be effective. To determine which DC subsets induce tolerance in the context of chronic autoimmunity, we used chimeric antibodies specific for DC inhibitory receptor 2 (DCIR2) or DEC-205 to target self-antigen to CD11b+ (cDC2) DCs and CD8+ (cDC1) DCs, respectively, in autoimmune-prone nonobese diabetic (NOD) mice. Antigen presentation by DCIR2+ DCs but not DEC-205+ DCs elicited tolerogenic CD4+ T-cell responses in NOD mice. β-Cell antigen delivered to DCIR2+ DCs delayed diabetes induction and induced increased T-cell apoptosis without interferon-γ (IFN-γ) or sustained expansion of autoreactive CD4+ T cells. These divergent responses were preceded by differential gene expression in T cells early after in vivo stimulation. Zbtb32 was higher in T cells stimulated with DCIR2+ DCs, and overexpression of Zbtb32 in T cells inhibited diabetes development, T-cell expansion, and IFN-γ production. Therefore, we have identified DCIR2+ DCs as capable of inducing antigen-specific tolerance in the face of ongoing autoimmunity and have also identified Zbtb32 as a suppressive transcription factor that controls T cell–mediated autoimmunity.

No MeSH data available.


Related in: MedlinePlus

DEC-205+ and DCIR2+ DCs induce similar Treg proliferation, with little Treg induction and no increase in the Treg-to-Teff ratio. A: BDC2.5 Foxp3+ T cells and BDC2.5 Foxp3− T cells, sorted from BDC2.5.Foxp3-GFP mice, were stimulated in vivo with αDEC-BDC or αDCIR2-BDC for 3 days in NOD mice. B: The number of GFP+ cells was assessed among transferred GFP+ cells after treatment with the indicated antibodies. Average of three independent experiments ± SEM. C: The number of GFP+ cells converted from transferred GFP− cells (left) and the number of GFP− cells (right) expanded by the indicated treatments were assessed. Average of three independent experiments ± SEM. D: NOD mice were injected with BDC2.5 T cells, and 5 days after treatment with the indicated antibodies, the total percentage of BDC2.5 T cells expressing Foxp3 was measured. Average of two independent experiments ± SEM. No significant differences were observed by statistical analysis with one-way ANOVA (B and C) or two-way ANOVA (D). LN, lymph nodes; ND, not detected; NS, not significant; pLN, pancreatic lymph nodes.
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Figure 5: DEC-205+ and DCIR2+ DCs induce similar Treg proliferation, with little Treg induction and no increase in the Treg-to-Teff ratio. A: BDC2.5 Foxp3+ T cells and BDC2.5 Foxp3− T cells, sorted from BDC2.5.Foxp3-GFP mice, were stimulated in vivo with αDEC-BDC or αDCIR2-BDC for 3 days in NOD mice. B: The number of GFP+ cells was assessed among transferred GFP+ cells after treatment with the indicated antibodies. Average of three independent experiments ± SEM. C: The number of GFP+ cells converted from transferred GFP− cells (left) and the number of GFP− cells (right) expanded by the indicated treatments were assessed. Average of three independent experiments ± SEM. D: NOD mice were injected with BDC2.5 T cells, and 5 days after treatment with the indicated antibodies, the total percentage of BDC2.5 T cells expressing Foxp3 was measured. Average of two independent experiments ± SEM. No significant differences were observed by statistical analysis with one-way ANOVA (B and C) or two-way ANOVA (D). LN, lymph nodes; ND, not detected; NS, not significant; pLN, pancreatic lymph nodes.

Mentions: DEC-205+ and DCIR2+ DC subsets have been reported to be important for Treg induction or expansion, respectively (34,41). To separately test induction or expansion of Tregs, GFP+ (Foxp3+) and GFP− CD4+ cells were sorted from NOD.BDC.Foxp3-GFP mice and injected into NOD mice along with DC-targeted antigen (Fig. 5A). Although αDEC-BDC and αDCIR2-BDC both caused proliferation and expansion of injected Foxp3+ Tregs at day 3, the expansion elicited by the two DC subsets was not significantly different, and this expansion was similar to the expansion of GFP− cells (Fig. 5B and C). Likewise, little conversion of GFP− to GFP+ cells was observed after stimulation by either DC subset (Fig. 5C). No significant increase in the percentage of Foxp3+ BDC2.5 T cells after stimulation of total CD4+ BDC2.5 T cells by DEC-205+ or DCIR2+ DCs was observed at antibody doses ranging from 10 to 1,000 ng (Fig. 5D). This suggests that Treg expansion is likely occurring at similar rates as parallel Teff responses. Therefore, Tregs do not likely contribute significantly to the differences in responses induced by DEC-205+ and DCIR2+ DCs, and neither DC subset in NOD mice is capable of increasing the Treg-to-Teff ratio to skew toward tolerance.


DCIR2 + cDC2 DCs and Zbtb32 Restore CD4 + T-Cell Tolerance and Inhibit Diabetes
DEC-205+ and DCIR2+ DCs induce similar Treg proliferation, with little Treg induction and no increase in the Treg-to-Teff ratio. A: BDC2.5 Foxp3+ T cells and BDC2.5 Foxp3− T cells, sorted from BDC2.5.Foxp3-GFP mice, were stimulated in vivo with αDEC-BDC or αDCIR2-BDC for 3 days in NOD mice. B: The number of GFP+ cells was assessed among transferred GFP+ cells after treatment with the indicated antibodies. Average of three independent experiments ± SEM. C: The number of GFP+ cells converted from transferred GFP− cells (left) and the number of GFP− cells (right) expanded by the indicated treatments were assessed. Average of three independent experiments ± SEM. D: NOD mice were injected with BDC2.5 T cells, and 5 days after treatment with the indicated antibodies, the total percentage of BDC2.5 T cells expressing Foxp3 was measured. Average of two independent experiments ± SEM. No significant differences were observed by statistical analysis with one-way ANOVA (B and C) or two-way ANOVA (D). LN, lymph nodes; ND, not detected; NS, not significant; pLN, pancreatic lymph nodes.
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Figure 5: DEC-205+ and DCIR2+ DCs induce similar Treg proliferation, with little Treg induction and no increase in the Treg-to-Teff ratio. A: BDC2.5 Foxp3+ T cells and BDC2.5 Foxp3− T cells, sorted from BDC2.5.Foxp3-GFP mice, were stimulated in vivo with αDEC-BDC or αDCIR2-BDC for 3 days in NOD mice. B: The number of GFP+ cells was assessed among transferred GFP+ cells after treatment with the indicated antibodies. Average of three independent experiments ± SEM. C: The number of GFP+ cells converted from transferred GFP− cells (left) and the number of GFP− cells (right) expanded by the indicated treatments were assessed. Average of three independent experiments ± SEM. D: NOD mice were injected with BDC2.5 T cells, and 5 days after treatment with the indicated antibodies, the total percentage of BDC2.5 T cells expressing Foxp3 was measured. Average of two independent experiments ± SEM. No significant differences were observed by statistical analysis with one-way ANOVA (B and C) or two-way ANOVA (D). LN, lymph nodes; ND, not detected; NS, not significant; pLN, pancreatic lymph nodes.
Mentions: DEC-205+ and DCIR2+ DC subsets have been reported to be important for Treg induction or expansion, respectively (34,41). To separately test induction or expansion of Tregs, GFP+ (Foxp3+) and GFP− CD4+ cells were sorted from NOD.BDC.Foxp3-GFP mice and injected into NOD mice along with DC-targeted antigen (Fig. 5A). Although αDEC-BDC and αDCIR2-BDC both caused proliferation and expansion of injected Foxp3+ Tregs at day 3, the expansion elicited by the two DC subsets was not significantly different, and this expansion was similar to the expansion of GFP− cells (Fig. 5B and C). Likewise, little conversion of GFP− to GFP+ cells was observed after stimulation by either DC subset (Fig. 5C). No significant increase in the percentage of Foxp3+ BDC2.5 T cells after stimulation of total CD4+ BDC2.5 T cells by DEC-205+ or DCIR2+ DCs was observed at antibody doses ranging from 10 to 1,000 ng (Fig. 5D). This suggests that Treg expansion is likely occurring at similar rates as parallel Teff responses. Therefore, Tregs do not likely contribute significantly to the differences in responses induced by DEC-205+ and DCIR2+ DCs, and neither DC subset in NOD mice is capable of increasing the Treg-to-Teff ratio to skew toward tolerance.

View Article: PubMed Central - PubMed

ABSTRACT

During autoimmunity, the normal ability of dendritic cells (DCs) to induce T-cell tolerance is disrupted; therefore, autoimmune disease therapies based on cell types and molecular pathways that elicit tolerance in the steady state may not be effective. To determine which DC subsets induce tolerance in the context of chronic autoimmunity, we used chimeric antibodies specific for DC inhibitory receptor 2 (DCIR2) or DEC-205 to target self-antigen to CD11b+ (cDC2) DCs and CD8+ (cDC1) DCs, respectively, in autoimmune-prone nonobese diabetic (NOD) mice. Antigen presentation by DCIR2+ DCs but not DEC-205+ DCs elicited tolerogenic CD4+ T-cell responses in NOD mice. β-Cell antigen delivered to DCIR2+ DCs delayed diabetes induction and induced increased T-cell apoptosis without interferon-γ (IFN-γ) or sustained expansion of autoreactive CD4+ T cells. These divergent responses were preceded by differential gene expression in T cells early after in vivo stimulation. Zbtb32 was higher in T cells stimulated with DCIR2+ DCs, and overexpression of Zbtb32 in T cells inhibited diabetes development, T-cell expansion, and IFN-γ production. Therefore, we have identified DCIR2+ DCs as capable of inducing antigen-specific tolerance in the face of ongoing autoimmunity and have also identified Zbtb32 as a suppressive transcription factor that controls T cell–mediated autoimmunity.

No MeSH data available.


Related in: MedlinePlus