Limits...
Interleukin-1β produced in response to islet autoantigen presentation differentiates T-helper 17 cells at the expense of regulatory T-cells: Implications for the timing of tolerizing immunotherapy.

Bertin-Maghit S, Pang D, O'Sullivan B, Best S, Duggan E, Paul S, Thomas H, Kay TW, Harrison LC, Steptoe R, Thomas R - Diabetes (2010)

Bottom Line: IL-1β production by NOD splenocytes and mRNA expression by islets increased from 6 to 16 weeks of age when major histocompatibility complex (MHC)-restricted islet antigen presentation to autoreactive T-cells occurred.IL-1 reduced the capacity of Treg cells to suppress effector cells and promoted their conversion to Th17 cells.RelB(lo) DCs exacerbated the IL-1-dependent decline in Treg function and promoted Th17 conversion.

View Article: PubMed Central - PubMed

Affiliation: The University of Queensland Diamantina Institute, Princess Alexandra Hospital, Brisbane, Australia.

ABSTRACT

Objective: The effectiveness of tolerizing immunotherapeutic strategies, such as anti-CD40L or dendritic cells (DCs), is greater when administered to young nonobese diabetic (NOD) mice than at peak insulitis. RelB(lo) DCs, generated in the presence of an nuclear factor-κB inhibitor, induce T-regulatory (Treg) cells and suppress inflammation in a model of rheumatoid arthritis. Interleukin (IL)-1β is overexpressed in humans and mice at risk of type 1 diabetes, dysregulates Treg cells, and accelerates diabetes in NOD mice. We investigated the relationship between IL-1β production and the response to RelB(lo) DCs in the prediabetic period.

Research design and methods: We injected RelB(lo) DCs subcutaneously into 4- or 14-week-old NOD mice and tracked the incidence of diabetes and effect on Treg cell function. We measured the expression of proinflammatory cytokines by stimulated splenocytes and unstimulated islets from mice of different ages and strains and proliferative and cytokine responses of T effectors to Treg in vitro.

Results: Tolerizing RelB(lo) DCs significantly inhibited diabetes progression when administered to 4-week-old but not 14-week-old mice. IL-1β production by NOD splenocytes and mRNA expression by islets increased from 6 to 16 weeks of age when major histocompatibility complex (MHC)-restricted islet antigen presentation to autoreactive T-cells occurred. IL-1 reduced the capacity of Treg cells to suppress effector cells and promoted their conversion to Th17 cells. RelB(lo) DCs exacerbated the IL-1-dependent decline in Treg function and promoted Th17 conversion.

Conclusions: IL-1β, generated by islet-autoreactive cells in MHC-susceptible mice, accelerates diabetes by differentiating Th17 at the expense of Treg. Tolerizing DC therapies can regulate islet autoantigen priming and prevent diabetes, but progression past the IL-1β/IL-17 checkpoint signals the need for other strategies.

Show MeSH

Related in: MedlinePlus

RelBlo tolerizing DCs inhibit diabetes when administered to young, but not insulitic, NOD mice. RelBlo DCs were generated from the bone marrow of NOD or pINS.NOD transgenic mice in the presence of GM-CSF, IL-4, and Bay11-7082. Control DCs were generated from the same mice in the presence of cytokines and the absence of Bay11-7082. Twenty-eight-day-old (A and B) or 100-day-old female NOD (C and D) mice were injected subcutaneously with 5 × 105 DCs. Mice were screened weekly for diabetes until 250 days of age. Diabetes incidence curves are shown for groups each containing 12 mice. *P < 0.05 (Kaplan-Meier survival analysis with Bonferroni correction for multiple groups). Insulitis was assessed at 12 weeks (E) in four female NOD mice per group treated at 4 weeks of age with DCs as shown. Data represent means ± SD.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3012178&req=5

Figure 1: RelBlo tolerizing DCs inhibit diabetes when administered to young, but not insulitic, NOD mice. RelBlo DCs were generated from the bone marrow of NOD or pINS.NOD transgenic mice in the presence of GM-CSF, IL-4, and Bay11-7082. Control DCs were generated from the same mice in the presence of cytokines and the absence of Bay11-7082. Twenty-eight-day-old (A and B) or 100-day-old female NOD (C and D) mice were injected subcutaneously with 5 × 105 DCs. Mice were screened weekly for diabetes until 250 days of age. Diabetes incidence curves are shown for groups each containing 12 mice. *P < 0.05 (Kaplan-Meier survival analysis with Bonferroni correction for multiple groups). Insulitis was assessed at 12 weeks (E) in four female NOD mice per group treated at 4 weeks of age with DCs as shown. Data represent means ± SD.

Mentions: To investigate the consequences of administration of RelBlo DCs to young or insulitic NOD mice, we generated RelBlo DCs from NOD mice by incubation of bone marrow cells in the presence of GM-CSF, IL-4, and Bay11-7082 (an irreversible inhibitor of NF-κB and inflammasome, but not p38 mitogen-activated protein kinase [40,41]) as previously described (33,35). CD40 and MHC class II expression were reduced by addition of Bay11-7082 to NOD DC cultures compared with control DCs without Bay11-7082, as expected (supplementary Fig. 1 in the online appendix, available at http://diabetes.diabetesjournals.org/cgi/content/full/db10-0104/DC1). The concentration of inhibitor required to inhibit expression of CD40 and MHC class II in NOD bone marrow cell cultures was generally 50% of that required for C57BL/6 RelBlo DC cultures, consistent with the constitutively higher levels of NF-κB expression in NOD cells (11). To determine the potential of RelBlo DCs to suppress disease generation in an antigen-specific manner, we also generated RelBlo DCs from NOD mice where proinsulin is driven as a transgene by the MHC class II promoter (pINS.NOD) (42). Mean survival without diabetes was increased from 161 days (95% CI 132–191) if mice were untreated to 232 days (215–248) in mice administered 5 × 105 RelBlo DCs subcutaneously, generated from NOD.Lt mice (P = 0.002). The smaller increases in survival time in mice administered pINS.NOD DCs, or control DCs, at 28 days of age were not statistically significant (Fig. 1). There was a trend for reduction in insulitis at 85 days in mice treated with NOD.Lt or pINS.NOD RelBlo DCs compared with PBS, but because islets from only four mice per group were examined, it was not meaningful to analyze the data statistically (Fig. 1E). In contrast, diabetes incidence was not reduced relative to saline-treated controls by transfer of RelBlo or control NOD or pINS.NOD DCs at 100 days of age and if anything, NOD RelBlo DC reduced the survival time without diabetes at this age (Fig. 1C and D). Thus, as expected, administration of RelBlo DCs prevented development of diabetes when administered to 28- but not 100-day-old NOD mice. This was not dependent on transgenic expression of proinsulin autoantigen by the DCs.


Interleukin-1β produced in response to islet autoantigen presentation differentiates T-helper 17 cells at the expense of regulatory T-cells: Implications for the timing of tolerizing immunotherapy.

Bertin-Maghit S, Pang D, O'Sullivan B, Best S, Duggan E, Paul S, Thomas H, Kay TW, Harrison LC, Steptoe R, Thomas R - Diabetes (2010)

RelBlo tolerizing DCs inhibit diabetes when administered to young, but not insulitic, NOD mice. RelBlo DCs were generated from the bone marrow of NOD or pINS.NOD transgenic mice in the presence of GM-CSF, IL-4, and Bay11-7082. Control DCs were generated from the same mice in the presence of cytokines and the absence of Bay11-7082. Twenty-eight-day-old (A and B) or 100-day-old female NOD (C and D) mice were injected subcutaneously with 5 × 105 DCs. Mice were screened weekly for diabetes until 250 days of age. Diabetes incidence curves are shown for groups each containing 12 mice. *P < 0.05 (Kaplan-Meier survival analysis with Bonferroni correction for multiple groups). Insulitis was assessed at 12 weeks (E) in four female NOD mice per group treated at 4 weeks of age with DCs as shown. Data represent means ± SD.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3012178&req=5

Figure 1: RelBlo tolerizing DCs inhibit diabetes when administered to young, but not insulitic, NOD mice. RelBlo DCs were generated from the bone marrow of NOD or pINS.NOD transgenic mice in the presence of GM-CSF, IL-4, and Bay11-7082. Control DCs were generated from the same mice in the presence of cytokines and the absence of Bay11-7082. Twenty-eight-day-old (A and B) or 100-day-old female NOD (C and D) mice were injected subcutaneously with 5 × 105 DCs. Mice were screened weekly for diabetes until 250 days of age. Diabetes incidence curves are shown for groups each containing 12 mice. *P < 0.05 (Kaplan-Meier survival analysis with Bonferroni correction for multiple groups). Insulitis was assessed at 12 weeks (E) in four female NOD mice per group treated at 4 weeks of age with DCs as shown. Data represent means ± SD.
Mentions: To investigate the consequences of administration of RelBlo DCs to young or insulitic NOD mice, we generated RelBlo DCs from NOD mice by incubation of bone marrow cells in the presence of GM-CSF, IL-4, and Bay11-7082 (an irreversible inhibitor of NF-κB and inflammasome, but not p38 mitogen-activated protein kinase [40,41]) as previously described (33,35). CD40 and MHC class II expression were reduced by addition of Bay11-7082 to NOD DC cultures compared with control DCs without Bay11-7082, as expected (supplementary Fig. 1 in the online appendix, available at http://diabetes.diabetesjournals.org/cgi/content/full/db10-0104/DC1). The concentration of inhibitor required to inhibit expression of CD40 and MHC class II in NOD bone marrow cell cultures was generally 50% of that required for C57BL/6 RelBlo DC cultures, consistent with the constitutively higher levels of NF-κB expression in NOD cells (11). To determine the potential of RelBlo DCs to suppress disease generation in an antigen-specific manner, we also generated RelBlo DCs from NOD mice where proinsulin is driven as a transgene by the MHC class II promoter (pINS.NOD) (42). Mean survival without diabetes was increased from 161 days (95% CI 132–191) if mice were untreated to 232 days (215–248) in mice administered 5 × 105 RelBlo DCs subcutaneously, generated from NOD.Lt mice (P = 0.002). The smaller increases in survival time in mice administered pINS.NOD DCs, or control DCs, at 28 days of age were not statistically significant (Fig. 1). There was a trend for reduction in insulitis at 85 days in mice treated with NOD.Lt or pINS.NOD RelBlo DCs compared with PBS, but because islets from only four mice per group were examined, it was not meaningful to analyze the data statistically (Fig. 1E). In contrast, diabetes incidence was not reduced relative to saline-treated controls by transfer of RelBlo or control NOD or pINS.NOD DCs at 100 days of age and if anything, NOD RelBlo DC reduced the survival time without diabetes at this age (Fig. 1C and D). Thus, as expected, administration of RelBlo DCs prevented development of diabetes when administered to 28- but not 100-day-old NOD mice. This was not dependent on transgenic expression of proinsulin autoantigen by the DCs.

Bottom Line: IL-1β production by NOD splenocytes and mRNA expression by islets increased from 6 to 16 weeks of age when major histocompatibility complex (MHC)-restricted islet antigen presentation to autoreactive T-cells occurred.IL-1 reduced the capacity of Treg cells to suppress effector cells and promoted their conversion to Th17 cells.RelB(lo) DCs exacerbated the IL-1-dependent decline in Treg function and promoted Th17 conversion.

View Article: PubMed Central - PubMed

Affiliation: The University of Queensland Diamantina Institute, Princess Alexandra Hospital, Brisbane, Australia.

ABSTRACT

Objective: The effectiveness of tolerizing immunotherapeutic strategies, such as anti-CD40L or dendritic cells (DCs), is greater when administered to young nonobese diabetic (NOD) mice than at peak insulitis. RelB(lo) DCs, generated in the presence of an nuclear factor-κB inhibitor, induce T-regulatory (Treg) cells and suppress inflammation in a model of rheumatoid arthritis. Interleukin (IL)-1β is overexpressed in humans and mice at risk of type 1 diabetes, dysregulates Treg cells, and accelerates diabetes in NOD mice. We investigated the relationship between IL-1β production and the response to RelB(lo) DCs in the prediabetic period.

Research design and methods: We injected RelB(lo) DCs subcutaneously into 4- or 14-week-old NOD mice and tracked the incidence of diabetes and effect on Treg cell function. We measured the expression of proinflammatory cytokines by stimulated splenocytes and unstimulated islets from mice of different ages and strains and proliferative and cytokine responses of T effectors to Treg in vitro.

Results: Tolerizing RelB(lo) DCs significantly inhibited diabetes progression when administered to 4-week-old but not 14-week-old mice. IL-1β production by NOD splenocytes and mRNA expression by islets increased from 6 to 16 weeks of age when major histocompatibility complex (MHC)-restricted islet antigen presentation to autoreactive T-cells occurred. IL-1 reduced the capacity of Treg cells to suppress effector cells and promoted their conversion to Th17 cells. RelB(lo) DCs exacerbated the IL-1-dependent decline in Treg function and promoted Th17 conversion.

Conclusions: IL-1β, generated by islet-autoreactive cells in MHC-susceptible mice, accelerates diabetes by differentiating Th17 at the expense of Treg. Tolerizing DC therapies can regulate islet autoantigen priming and prevent diabetes, but progression past the IL-1β/IL-17 checkpoint signals the need for other strategies.

Show MeSH
Related in: MedlinePlus