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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-22andIL-22receptor(IL-22Rα) in the pancreatic islet cells after CFA immunization. The relative mRNA expression levels of IL-22 and IL-22Rα (A and B) was carried out in the pancreas of non-diabetic (4-week old), pre-diabetic (8-week old and 12-week old), and older (>16 week-old) diabetic NOD mice following CFA immunization. Female NOD mice were injected i.p. with either 100 μl of CFA emulsified in saline or with saline alone and 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 and age group have been compared to 4-wk-old saline-treated mice and represent the average fold change of mRNA expression ± SEM. The relative expression of mRNA was taken from three pooled samples of RNA per group (with three mice per pooled sample). Age groups indicated by a double asterisk (**) are significantly different (P<0.05); treatments indicated by the asterisk (*) are significantly different from the saline control (P<0.05).
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Fig2: Expression ofIL-22andIL-22receptor(IL-22Rα) in the pancreatic islet cells after CFA immunization. The relative mRNA expression levels of IL-22 and IL-22Rα (A and B) was carried out in the pancreas of non-diabetic (4-week old), pre-diabetic (8-week old and 12-week old), and older (>16 week-old) diabetic NOD mice following CFA immunization. Female NOD mice were injected i.p. with either 100 μl of CFA emulsified in saline or with saline alone and 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 and age group have been compared to 4-wk-old saline-treated mice and represent the average fold change of mRNA expression ± SEM. The relative expression of mRNA was taken from three pooled samples of RNA per group (with three mice per pooled sample). Age groups indicated by a double asterisk (**) are significantly different (P<0.05); treatments indicated by the asterisk (*) are significantly different from the saline control (P<0.05).

Mentions: We have previously shown that CFA injection in non-diabetic and diabetic NOD mice results in a substantial increase in Reg gene expression [5]. To determine whether the development of diabetes was accompanied by similar changes in IL-22 gene expression in vivo, qRT-PCR analysis was performed using IL-22 gene-specific primers for cDNA using the various NOD mouse treatment/age groups. As shown in Figure 2A a significant effect of age on IL-22 expression was detected (F= 6.374; df= 3; P=0.021). Post-hoc analysis revealed IL-22 mRNA abundance in the pancreas to be significantly higher in diabetic mice when compared to non-diabetic (4-week-old) animals (P<0.05). The mean IL-22 expression tended to be higher in the pre-diabetic (8-week-old) mice and in diabetic mice following CFA treatment (by approximately 2 and 4-fold respectively).Figure 2


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-22andIL-22receptor(IL-22Rα) in the pancreatic islet cells after CFA immunization. The relative mRNA expression levels of IL-22 and IL-22Rα (A and B) was carried out in the pancreas of non-diabetic (4-week old), pre-diabetic (8-week old and 12-week old), and older (>16 week-old) diabetic NOD mice following CFA immunization. Female NOD mice were injected i.p. with either 100 μl of CFA emulsified in saline or with saline alone and 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 and age group have been compared to 4-wk-old saline-treated mice and represent the average fold change of mRNA expression ± SEM. The relative expression of mRNA was taken from three pooled samples of RNA per group (with three mice per pooled sample). Age groups indicated by a double asterisk (**) are significantly different (P<0.05); treatments indicated by the asterisk (*) are significantly different 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

Fig2: Expression ofIL-22andIL-22receptor(IL-22Rα) in the pancreatic islet cells after CFA immunization. The relative mRNA expression levels of IL-22 and IL-22Rα (A and B) was carried out in the pancreas of non-diabetic (4-week old), pre-diabetic (8-week old and 12-week old), and older (>16 week-old) diabetic NOD mice following CFA immunization. Female NOD mice were injected i.p. with either 100 μl of CFA emulsified in saline or with saline alone and 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 and age group have been compared to 4-wk-old saline-treated mice and represent the average fold change of mRNA expression ± SEM. The relative expression of mRNA was taken from three pooled samples of RNA per group (with three mice per pooled sample). Age groups indicated by a double asterisk (**) are significantly different (P<0.05); treatments indicated by the asterisk (*) are significantly different from the saline control (P<0.05).
Mentions: We have previously shown that CFA injection in non-diabetic and diabetic NOD mice results in a substantial increase in Reg gene expression [5]. To determine whether the development of diabetes was accompanied by similar changes in IL-22 gene expression in vivo, qRT-PCR analysis was performed using IL-22 gene-specific primers for cDNA using the various NOD mouse treatment/age groups. As shown in Figure 2A a significant effect of age on IL-22 expression was detected (F= 6.374; df= 3; P=0.021). Post-hoc analysis revealed IL-22 mRNA abundance in the pancreas to be significantly higher in diabetic mice when compared to non-diabetic (4-week-old) animals (P<0.05). The mean IL-22 expression tended to be higher in the pre-diabetic (8-week-old) mice and in diabetic mice following CFA treatment (by approximately 2 and 4-fold respectively).Figure 2

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