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CD4+CD25+ T regulatory cells dependent on ICOS promote regulation of effector cells in the prediabetic lesion.

Herman AE, Freeman GJ, Mathis D, Benoist C - J. Exp. Med. (2004)

Bottom Line: Here, we report that Tregs and T effector cells (Teffs) coexist within the pancreatic lesion before type 1 diabetes onset.We find that BDC2.5 T cell receptor transgenic animals contain a small subset of FoxP3 positive CD4+CD25+CD69- cells in the pancreas, actively turning over, expressing the clonotypic receptor, and containing functional regulatory activity.Blockade of ICOS rapidly converts early insulitis to diabetes, which disrupts the balance of Teffs and Tregs and promotes a very broad shift in the expression of the T regulatory-specific profile.

View Article: PubMed Central - PubMed

Affiliation: Section on Immunology and Immunogenetics, Joslin Diabetes Center, Boston, MA 02215, USA.

ABSTRACT
CD4+CD25+ T regulatory cells (Tregs) prevent autoimmune disease, yet little is known about precisely where they exert their influence naturally in a spontaneous autoimmune disorder. Here, we report that Tregs and T effector cells (Teffs) coexist within the pancreatic lesion before type 1 diabetes onset. We find that BDC2.5 T cell receptor transgenic animals contain a small subset of FoxP3 positive CD4+CD25+CD69- cells in the pancreas, actively turning over, expressing the clonotypic receptor, and containing functional regulatory activity. Gene expression profiling confirms that the CD4+CD25+CD69- cells in pancreatic tissue express transcripts diagnostic of regulatory cells, but with significantly higher levels of interleukin 10 and inducible costimulator (ICOS) than their lymph node counterparts. Blockade of ICOS rapidly converts early insulitis to diabetes, which disrupts the balance of Teffs and Tregs and promotes a very broad shift in the expression of the T regulatory-specific profile. Thus, CD4+CD25+69- Tregs operate directly in the autoimmune lesion and are dependent on ICOS to keep it in a nondestructive state.

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ICOS blockade dampens the immunoregulatory gene profile and correlates with expression of proinflammatory cytokines. (A) Cells from pancreatic infiltrate of 3–4-wk-old BDC2.5 mice were isolated after treatment with anti-ICOS mAb or rat IgG control as indicated, and analyzed for CD25/CD69 expression profiles on CD4+B220− live lymphocytes. Data represent five separate experiments. Cells were sorted to high purity directly from the pancreatic lesion of 3–4-wk-old BDC2.5 mice 5 d after indicated treatments for the CD4+CD25+CD69− Treg population, or the CD25loCD69+ and CD25−CD69− combined Teff populations. RNA was prepared and hybridized to Affymetrix U74Av2 array GeneChips®, and data were analyzed as described in Materials and Methods and in Fig. 4. (B) Expression value comparison plot for Teff populations with anti-ICOS versus control treatment. (C) Similar plot for Treg populations. (D) A ratio plot comparing the ratio of Treg anti-ICOS–treated/Teff anti-ICOS–treated profiles to the same ratio in the Treg and Teff control-treated profiles. Diagonal line indicates where points would fall if nothing changed between the treatments. A selection of genes that are highly over- or underexpressed after anti-ICOS treatment are highlighted. (E) Changes in expression values for cytokines within the lesion in Treg and Teff cells from each treatment group are shown. Asterisks indicate fold changes >1.9.
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fig7: ICOS blockade dampens the immunoregulatory gene profile and correlates with expression of proinflammatory cytokines. (A) Cells from pancreatic infiltrate of 3–4-wk-old BDC2.5 mice were isolated after treatment with anti-ICOS mAb or rat IgG control as indicated, and analyzed for CD25/CD69 expression profiles on CD4+B220− live lymphocytes. Data represent five separate experiments. Cells were sorted to high purity directly from the pancreatic lesion of 3–4-wk-old BDC2.5 mice 5 d after indicated treatments for the CD4+CD25+CD69− Treg population, or the CD25loCD69+ and CD25−CD69− combined Teff populations. RNA was prepared and hybridized to Affymetrix U74Av2 array GeneChips®, and data were analyzed as described in Materials and Methods and in Fig. 4. (B) Expression value comparison plot for Teff populations with anti-ICOS versus control treatment. (C) Similar plot for Treg populations. (D) A ratio plot comparing the ratio of Treg anti-ICOS–treated/Teff anti-ICOS–treated profiles to the same ratio in the Treg and Teff control-treated profiles. Diagonal line indicates where points would fall if nothing changed between the treatments. A selection of genes that are highly over- or underexpressed after anti-ICOS treatment are highlighted. (E) Changes in expression values for cytokines within the lesion in Treg and Teff cells from each treatment group are shown. Asterisks indicate fold changes >1.9.

Mentions: Microarrays were used to analyze gene expression changes induced by ICOS blockade in the two populations. Teff or Treg CD4+ subsets were sorted as aforementioned from 4-wk-old BDC2.5/NOD mice 5 d after treatment with anti-ICOS or control rat IgG (Fig. 7 A), and RNA probes were prepared for microarray analysis as aforementioned. Changes induced by ICOS blockade were more limited in the effector population than in Tregs; the comparative plot was much tighter for Teff than for Tregs (Fig. 7, B and C). In Teff, only 33 genes were induced by a factor of ≥1.5-fold, and only 1 was repressed. Interestingly, a number of these induced genes have also been found activated in a recent analysis of pancreatic destruction induced by cyclophosphamide treatment of BDC2.5/NOD mice, including IFN-γ and several IFN-γ–regulated genes (Matos, M., personal communication).


CD4+CD25+ T regulatory cells dependent on ICOS promote regulation of effector cells in the prediabetic lesion.

Herman AE, Freeman GJ, Mathis D, Benoist C - J. Exp. Med. (2004)

ICOS blockade dampens the immunoregulatory gene profile and correlates with expression of proinflammatory cytokines. (A) Cells from pancreatic infiltrate of 3–4-wk-old BDC2.5 mice were isolated after treatment with anti-ICOS mAb or rat IgG control as indicated, and analyzed for CD25/CD69 expression profiles on CD4+B220− live lymphocytes. Data represent five separate experiments. Cells were sorted to high purity directly from the pancreatic lesion of 3–4-wk-old BDC2.5 mice 5 d after indicated treatments for the CD4+CD25+CD69− Treg population, or the CD25loCD69+ and CD25−CD69− combined Teff populations. RNA was prepared and hybridized to Affymetrix U74Av2 array GeneChips®, and data were analyzed as described in Materials and Methods and in Fig. 4. (B) Expression value comparison plot for Teff populations with anti-ICOS versus control treatment. (C) Similar plot for Treg populations. (D) A ratio plot comparing the ratio of Treg anti-ICOS–treated/Teff anti-ICOS–treated profiles to the same ratio in the Treg and Teff control-treated profiles. Diagonal line indicates where points would fall if nothing changed between the treatments. A selection of genes that are highly over- or underexpressed after anti-ICOS treatment are highlighted. (E) Changes in expression values for cytokines within the lesion in Treg and Teff cells from each treatment group are shown. Asterisks indicate fold changes >1.9.
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Related In: Results  -  Collection

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fig7: ICOS blockade dampens the immunoregulatory gene profile and correlates with expression of proinflammatory cytokines. (A) Cells from pancreatic infiltrate of 3–4-wk-old BDC2.5 mice were isolated after treatment with anti-ICOS mAb or rat IgG control as indicated, and analyzed for CD25/CD69 expression profiles on CD4+B220− live lymphocytes. Data represent five separate experiments. Cells were sorted to high purity directly from the pancreatic lesion of 3–4-wk-old BDC2.5 mice 5 d after indicated treatments for the CD4+CD25+CD69− Treg population, or the CD25loCD69+ and CD25−CD69− combined Teff populations. RNA was prepared and hybridized to Affymetrix U74Av2 array GeneChips®, and data were analyzed as described in Materials and Methods and in Fig. 4. (B) Expression value comparison plot for Teff populations with anti-ICOS versus control treatment. (C) Similar plot for Treg populations. (D) A ratio plot comparing the ratio of Treg anti-ICOS–treated/Teff anti-ICOS–treated profiles to the same ratio in the Treg and Teff control-treated profiles. Diagonal line indicates where points would fall if nothing changed between the treatments. A selection of genes that are highly over- or underexpressed after anti-ICOS treatment are highlighted. (E) Changes in expression values for cytokines within the lesion in Treg and Teff cells from each treatment group are shown. Asterisks indicate fold changes >1.9.
Mentions: Microarrays were used to analyze gene expression changes induced by ICOS blockade in the two populations. Teff or Treg CD4+ subsets were sorted as aforementioned from 4-wk-old BDC2.5/NOD mice 5 d after treatment with anti-ICOS or control rat IgG (Fig. 7 A), and RNA probes were prepared for microarray analysis as aforementioned. Changes induced by ICOS blockade were more limited in the effector population than in Tregs; the comparative plot was much tighter for Teff than for Tregs (Fig. 7, B and C). In Teff, only 33 genes were induced by a factor of ≥1.5-fold, and only 1 was repressed. Interestingly, a number of these induced genes have also been found activated in a recent analysis of pancreatic destruction induced by cyclophosphamide treatment of BDC2.5/NOD mice, including IFN-γ and several IFN-γ–regulated genes (Matos, M., personal communication).

Bottom Line: Here, we report that Tregs and T effector cells (Teffs) coexist within the pancreatic lesion before type 1 diabetes onset.We find that BDC2.5 T cell receptor transgenic animals contain a small subset of FoxP3 positive CD4+CD25+CD69- cells in the pancreas, actively turning over, expressing the clonotypic receptor, and containing functional regulatory activity.Blockade of ICOS rapidly converts early insulitis to diabetes, which disrupts the balance of Teffs and Tregs and promotes a very broad shift in the expression of the T regulatory-specific profile.

View Article: PubMed Central - PubMed

Affiliation: Section on Immunology and Immunogenetics, Joslin Diabetes Center, Boston, MA 02215, USA.

ABSTRACT
CD4+CD25+ T regulatory cells (Tregs) prevent autoimmune disease, yet little is known about precisely where they exert their influence naturally in a spontaneous autoimmune disorder. Here, we report that Tregs and T effector cells (Teffs) coexist within the pancreatic lesion before type 1 diabetes onset. We find that BDC2.5 T cell receptor transgenic animals contain a small subset of FoxP3 positive CD4+CD25+CD69- cells in the pancreas, actively turning over, expressing the clonotypic receptor, and containing functional regulatory activity. Gene expression profiling confirms that the CD4+CD25+CD69- cells in pancreatic tissue express transcripts diagnostic of regulatory cells, but with significantly higher levels of interleukin 10 and inducible costimulator (ICOS) than their lymph node counterparts. Blockade of ICOS rapidly converts early insulitis to diabetes, which disrupts the balance of Teffs and Tregs and promotes a very broad shift in the expression of the T regulatory-specific profile. Thus, CD4+CD25+69- Tregs operate directly in the autoimmune lesion and are dependent on ICOS to keep it in a nondestructive state.

Show MeSH
Related in: MedlinePlus