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The involvement of interleukin-22 in the expression of pancreatic beta cell regenerative Reg genes.

Hill T, Krougly O, Nikoopour E, Bellemore S, Lee-Chan E, Fouser LA, Hill DJ, Singh B - Cell Regen (Lond) (2013)

Bottom Line: This was associated with increased islet Regenerating (Reg) genes expression, and elevated IL-22-producing Th17 T-cells in the pancreas.Our results showed: 1) Reg1 and Reg2 mRNA abundance to be significantly increased in IL-22-treated islets in vitro; 2) IL-22 mRNA expression in the pre-diabetic mouse pancreas increased with time following CFA treatment; 3) a reduced expression of IL-22Rα following CFA treatment; 4) a down-regulation in Reg1 and Reg2 mRNA expression in the pancreas of pre-diabetic mice injected with an IL-22 neutralizing antibody; and 5) an increased islet β-cell DNA synthesis in vitro in the presence of IL-22.We conclude that IL-22 may contribute to the regeneration of β-cells by up-regulating Regenerating Reg1 and Reg2 genes in the islets.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, University of Western Ontario, London, ON Canada.

ABSTRACT

Background: In Type 1 diabetes, the insulin-producing β-cells within the pancreatic islets of Langerhans are destroyed. We showed previously that immunotherapy with Bacillus Calmette-Guerin (BCG) or complete Freund's adjuvant (CFA) of non-obese diabetic (NOD) mice can prevent disease process and pancreatic β-cell loss. This was associated with increased islet Regenerating (Reg) genes expression, and elevated IL-22-producing Th17 T-cells in the pancreas.

Results: We hypothesized that IL-22 was responsible for the increased Reg gene expression in the pancreas. We therefore quantified the Reg1, Reg2, and Reg3δ (INGAP) mRNA expression in isolated pre-diabetic NOD islets treated with IL-22. We measured IL-22, and IL-22 receptor(R)-α mRNA expression in the pancreas and spleen of pre-diabetic and diabetic NOD mice. Our results showed: 1) Reg1 and Reg2 mRNA abundance to be significantly increased in IL-22-treated islets in vitro; 2) IL-22 mRNA expression in the pre-diabetic mouse pancreas increased with time following CFA treatment; 3) a reduced expression of IL-22Rα following CFA treatment; 4) a down-regulation in Reg1 and Reg2 mRNA expression in the pancreas of pre-diabetic mice injected with an IL-22 neutralizing antibody; and 5) an increased islet β-cell DNA synthesis in vitro in the presence of IL-22.

Conclusions: We conclude that IL-22 may contribute to the regeneration of β-cells by up-regulating Regenerating Reg1 and Reg2 genes in the islets.

No MeSH data available.


Related in: MedlinePlus

Expression of Reg genes in the pancreatic islet cells after IL-22 neutralization.Reg1, Reg2, and Reg3δ gene expression levels (A-C) was determined in the pancreas of 6-week-old NOD mice immunized with CFA and treated with an IL-22 neutralizing or a control antibody. Female NOD mice were injected i.p. with either 100 μl of CFA emulsified in saline followed by 100 μl of IL-22 neutralizing antibody (IL-22-01) 1 hr later, or 100 μl of CFA emulsified in saline followed by 100 μl of isotype control antibody. Mice were sacrificed 48 hrs following immunization. Whole pancreatic tissue was homogenized and total RNA extracted for reverse transcription quantitative real-time PCR analysis using gene-specific primers. The results shown for each treatment have been compared to mice immunized with CFA and the IgG isotype antibody, and represent the average fold change of mRNA expression ± SEM. The relative expression of mRNA was taken from three pooled samples per group (with three mice per pooled sample). Treatments indicated by the asterisk (*) are significantly different from the CFA control (P<0.001).
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Fig3: Expression of Reg genes in the pancreatic islet cells after IL-22 neutralization.Reg1, Reg2, and Reg3δ gene expression levels (A-C) was determined in the pancreas of 6-week-old NOD mice immunized with CFA and treated with an IL-22 neutralizing or a control antibody. Female NOD mice were injected i.p. with either 100 μl of CFA emulsified in saline followed by 100 μl of IL-22 neutralizing antibody (IL-22-01) 1 hr later, or 100 μl of CFA emulsified in saline followed by 100 μl of isotype control antibody. Mice were sacrificed 48 hrs following immunization. Whole pancreatic tissue was homogenized and total RNA extracted for reverse transcription quantitative real-time PCR analysis using gene-specific primers. The results shown for each treatment have been compared to mice immunized with CFA and the IgG isotype antibody, and represent the average fold change of mRNA expression ± SEM. The relative expression of mRNA was taken from three pooled samples per group (with three mice per pooled sample). Treatments indicated by the asterisk (*) are significantly different from the CFA control (P<0.001).

Mentions: To determine whether IL-22 was directly involved in the up-regulation of Reg expression following CFA treatment, the abundance of Reg2, Reg1and Reg3δ mRNAs were measured in the pancreas of 6-week-old NOD mice immunized with CFA followed by i.p injection of an IL-22 neutralizing antibody 1 h later. Mice immunized with CFA and treated with the IL-22 neutralizing antibody [21] were found to have a significant reduction (approximately 480-fold) in Reg2 gene expression when compared to mice immunized with CFA and treated with an isotype control antibody (P<0.001) (Figure 3A). Likewise, Reg1 gene expression was significantly down-regulated in mice injected with the IL-22 neutralizing antibody (P<0.001) (Figure 3B). In contrast, however, no change in Reg3δ expression was observed for mice immunized with CFA in the presence of IL-22 neutralizing antibody when compared to mice injected with the isotype control antibody (Figure 3C).Figure 3


The involvement of interleukin-22 in the expression of pancreatic beta cell regenerative Reg genes.

Hill T, Krougly O, Nikoopour E, Bellemore S, Lee-Chan E, Fouser LA, Hill DJ, Singh B - Cell Regen (Lond) (2013)

Expression of Reg genes in the pancreatic islet cells after IL-22 neutralization.Reg1, Reg2, and Reg3δ gene expression levels (A-C) was determined in the pancreas of 6-week-old NOD mice immunized with CFA and treated with an IL-22 neutralizing or a control antibody. Female NOD mice were injected i.p. with either 100 μl of CFA emulsified in saline followed by 100 μl of IL-22 neutralizing antibody (IL-22-01) 1 hr later, or 100 μl of CFA emulsified in saline followed by 100 μl of isotype control antibody. Mice were sacrificed 48 hrs following immunization. Whole pancreatic tissue was homogenized and total RNA extracted for reverse transcription quantitative real-time PCR analysis using gene-specific primers. The results shown for each treatment have been compared to mice immunized with CFA and the IgG isotype antibody, and represent the average fold change of mRNA expression ± SEM. The relative expression of mRNA was taken from three pooled samples per group (with three mice per pooled sample). Treatments indicated by the asterisk (*) are significantly different from the CFA control (P<0.001).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4230743&req=5

Fig3: Expression of Reg genes in the pancreatic islet cells after IL-22 neutralization.Reg1, Reg2, and Reg3δ gene expression levels (A-C) was determined in the pancreas of 6-week-old NOD mice immunized with CFA and treated with an IL-22 neutralizing or a control antibody. Female NOD mice were injected i.p. with either 100 μl of CFA emulsified in saline followed by 100 μl of IL-22 neutralizing antibody (IL-22-01) 1 hr later, or 100 μl of CFA emulsified in saline followed by 100 μl of isotype control antibody. Mice were sacrificed 48 hrs following immunization. Whole pancreatic tissue was homogenized and total RNA extracted for reverse transcription quantitative real-time PCR analysis using gene-specific primers. The results shown for each treatment have been compared to mice immunized with CFA and the IgG isotype antibody, and represent the average fold change of mRNA expression ± SEM. The relative expression of mRNA was taken from three pooled samples per group (with three mice per pooled sample). Treatments indicated by the asterisk (*) are significantly different from the CFA control (P<0.001).
Mentions: To determine whether IL-22 was directly involved in the up-regulation of Reg expression following CFA treatment, the abundance of Reg2, Reg1and Reg3δ mRNAs were measured in the pancreas of 6-week-old NOD mice immunized with CFA followed by i.p injection of an IL-22 neutralizing antibody 1 h later. Mice immunized with CFA and treated with the IL-22 neutralizing antibody [21] were found to have a significant reduction (approximately 480-fold) in Reg2 gene expression when compared to mice immunized with CFA and treated with an isotype control antibody (P<0.001) (Figure 3A). Likewise, Reg1 gene expression was significantly down-regulated in mice injected with the IL-22 neutralizing antibody (P<0.001) (Figure 3B). In contrast, however, no change in Reg3δ expression was observed for mice immunized with CFA in the presence of IL-22 neutralizing antibody when compared to mice injected with the isotype control antibody (Figure 3C).Figure 3

Bottom Line: This was associated with increased islet Regenerating (Reg) genes expression, and elevated IL-22-producing Th17 T-cells in the pancreas.Our results showed: 1) Reg1 and Reg2 mRNA abundance to be significantly increased in IL-22-treated islets in vitro; 2) IL-22 mRNA expression in the pre-diabetic mouse pancreas increased with time following CFA treatment; 3) a reduced expression of IL-22Rα following CFA treatment; 4) a down-regulation in Reg1 and Reg2 mRNA expression in the pancreas of pre-diabetic mice injected with an IL-22 neutralizing antibody; and 5) an increased islet β-cell DNA synthesis in vitro in the presence of IL-22.We conclude that IL-22 may contribute to the regeneration of β-cells by up-regulating Regenerating Reg1 and Reg2 genes in the islets.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, University of Western Ontario, London, ON Canada.

ABSTRACT

Background: In Type 1 diabetes, the insulin-producing β-cells within the pancreatic islets of Langerhans are destroyed. We showed previously that immunotherapy with Bacillus Calmette-Guerin (BCG) or complete Freund's adjuvant (CFA) of non-obese diabetic (NOD) mice can prevent disease process and pancreatic β-cell loss. This was associated with increased islet Regenerating (Reg) genes expression, and elevated IL-22-producing Th17 T-cells in the pancreas.

Results: We hypothesized that IL-22 was responsible for the increased Reg gene expression in the pancreas. We therefore quantified the Reg1, Reg2, and Reg3δ (INGAP) mRNA expression in isolated pre-diabetic NOD islets treated with IL-22. We measured IL-22, and IL-22 receptor(R)-α mRNA expression in the pancreas and spleen of pre-diabetic and diabetic NOD mice. Our results showed: 1) Reg1 and Reg2 mRNA abundance to be significantly increased in IL-22-treated islets in vitro; 2) IL-22 mRNA expression in the pre-diabetic mouse pancreas increased with time following CFA treatment; 3) a reduced expression of IL-22Rα following CFA treatment; 4) a down-regulation in Reg1 and Reg2 mRNA expression in the pancreas of pre-diabetic mice injected with an IL-22 neutralizing antibody; and 5) an increased islet β-cell DNA synthesis in vitro in the presence of IL-22.

Conclusions: We conclude that IL-22 may contribute to the regeneration of β-cells by up-regulating Regenerating Reg1 and Reg2 genes in the islets.

No MeSH data available.


Related in: MedlinePlus