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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 ofIL-22in the spleen of 4-week-old non-diabetic NOD mice following CFA-treatment. The relative mRNA expression of IL-22 in the spleen of 4-week-old NOD mice was carried out following CFA-treatment. Quantitative RT-PCR analysis was performed using gene-specific primers (Table 1) on total RNA isolated from whole splenic tissue from female NOD mice injected i.p. with either 100 μl of CFA emulsified in saline or saline alone. Results shown represent the average fold-change in mRNA expression ± SEM compared with saline-treated mice. Results were taken from 3 mice per treatment. The asterisk (*) denotes a significant difference from the saline control (P<0.05).
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Fig4: Expression ofIL-22in the spleen of 4-week-old non-diabetic NOD mice following CFA-treatment. The relative mRNA expression of IL-22 in the spleen of 4-week-old NOD mice was carried out following CFA-treatment. Quantitative RT-PCR analysis was performed using gene-specific primers (Table 1) on total RNA isolated from whole splenic tissue from female NOD mice injected i.p. with either 100 μl of CFA emulsified in saline or saline alone. Results shown represent the average fold-change in mRNA expression ± SEM compared with saline-treated mice. Results were taken from 3 mice per treatment. The asterisk (*) denotes a significant difference from the saline control (P<0.05).

Mentions: Since the spleen is a major peripheral lymphoid tissue, we performed quantitative RT-PCR analysis on splenocytes derived from 4-week-old, pre-diabetic NOD mice, with or without CFA treatment, to investigate the presence of IL-22-producing Th17 cells during autoimmune insulitis (Figure 4). CFA treatment was found to significantly up-regulate IL-22 expression in the non-diabetic, 4 week-old mice by approximately 12-fold (P<0.05). To determine whether CFA-mediated Reg gene up-regulation occurred in the spleen as well as in pancreatic tissue, Reg1, Reg2 and Reg3δ mRNA expression was also analyzed in whole splenic tissue using non-diabetic mice with/without CFA injection. Similar to Huszarik et al.[5], no effect of treatment was detected on Reg mRNA expression (data not shown).Figure 4


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 ofIL-22in the spleen of 4-week-old non-diabetic NOD mice following CFA-treatment. The relative mRNA expression of IL-22 in the spleen of 4-week-old NOD mice was carried out following CFA-treatment. Quantitative RT-PCR analysis was performed using gene-specific primers (Table 1) on total RNA isolated from whole splenic tissue from female NOD mice injected i.p. with either 100 μl of CFA emulsified in saline or saline alone. Results shown represent the average fold-change in mRNA expression ± SEM compared with saline-treated mice. Results were taken from 3 mice per treatment. The asterisk (*) denotes a significant difference from the saline control (P<0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig4: Expression ofIL-22in the spleen of 4-week-old non-diabetic NOD mice following CFA-treatment. The relative mRNA expression of IL-22 in the spleen of 4-week-old NOD mice was carried out following CFA-treatment. Quantitative RT-PCR analysis was performed using gene-specific primers (Table 1) on total RNA isolated from whole splenic tissue from female NOD mice injected i.p. with either 100 μl of CFA emulsified in saline or saline alone. Results shown represent the average fold-change in mRNA expression ± SEM compared with saline-treated mice. Results were taken from 3 mice per treatment. The asterisk (*) denotes a significant difference from the saline control (P<0.05).
Mentions: Since the spleen is a major peripheral lymphoid tissue, we performed quantitative RT-PCR analysis on splenocytes derived from 4-week-old, pre-diabetic NOD mice, with or without CFA treatment, to investigate the presence of IL-22-producing Th17 cells during autoimmune insulitis (Figure 4). CFA treatment was found to significantly up-regulate IL-22 expression in the non-diabetic, 4 week-old mice by approximately 12-fold (P<0.05). To determine whether CFA-mediated Reg gene up-regulation occurred in the spleen as well as in pancreatic tissue, Reg1, Reg2 and Reg3δ mRNA expression was also analyzed in whole splenic tissue using non-diabetic mice with/without CFA injection. Similar to Huszarik et al.[5], no effect of treatment was detected on Reg mRNA expression (data not shown).Figure 4

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