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The expression of the beta cell-derived autoimmune ligand for the killer receptor nkp46 is attenuated in type 2 diabetes.

Gur C, Enk J, Weitman E, Bachar E, Suissa Y, Cohen G, Schyr RB, Sabanay H, Horwitz E, Glaser B, Dor Y, Pribluda A, Hanna JH, Leibowitz G, Mandelboim O - PLoS ONE (2013)

Bottom Line: Following glucose stimulation the NKp46 ligand translocates to the cell membrane and its secretion decreases.We further demonstrate by using several modalities that the unknown NKp46 ligand is not insulin.We demonstrate that the expression of the NKp46 ligand is decreased in all models of T2D studied, suggesting that NKp46 is not involved in T2D.

View Article: PubMed Central - PubMed

Affiliation: The Lautenberg Center for General and Tumor Immunology, The Department of Immunology and Cancer Research, The Institute for Medical Research Israel-Canada, The Hebrew University-Hadassah Medical School, Jerusalem, Israel.

ABSTRACT
NK cells rapidly kill tumor cells, virus infected cells and even self cells. This is mediated via killer receptors, among which NKp46 (NCR1 in mice) is prominent. We have recently demonstrated that in type 1 diabetes (T1D) NK cells accumulate in the diseased pancreas and that they manifest a hyporesponsive phenotype. In addition, we found that NKp46 recognizes an unknown ligand expressed by beta cells derived from humans and mice and that blocking of NKp46 activity prevented diabetes development. Here we investigated the properties of the unknown NKp46 ligand. We show that the NKp46 ligand is mainly located in insulin granules and that it is constitutively secreted. Following glucose stimulation the NKp46 ligand translocates to the cell membrane and its secretion decreases. We further demonstrate by using several modalities that the unknown NKp46 ligand is not insulin. Finally, we studied the expression of the NKp46 ligand in type 2 diabetes (T2D) using 3 different in vivo models and 2 species; mice and gerbils. We demonstrate that the expression of the NKp46 ligand is decreased in all models of T2D studied, suggesting that NKp46 is not involved in T2D.

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Expression of the NKp46 ligand in beta cells derived from P. Obesus.(A)Immunofluorescence staining of pancreatic tissues derived from 8–10 weeks of age, male, P. obesus gerbils fed with LE diet. In the upper images staining for insulin and for NKp46 ligand are shown and in the lower images staining for NKp46 ligand and glucagon are shown. (B) NKp46-Ig and anti-insulin staining of pancreases derived from diabetic (non-fasting blood glucose>250 mg/dl for 2 sequential days) P. obesus gerbils fed with HE diet for 21–24 days. (C) NKp46-Ig and anti-insulin staining of pancreatic tissues derived from diabetic P. obesus gerbils fed with HE diet for 21–24 days followed by 48 hours of LE diet (C, upper) or overnight (O/N) fasting (C, lower). (D) NKp46-Ig and anti-insulin staining of pancreatic tissues derived from non-diabetic P. obesus fed for 21–24 days with HE diet and then injected with Exendin 4 (EX4). For all Figure parts the left images represent staining with NKp46-Ig (NKp46 ligand, green), the middle images represent staining with anti-insulin (A, upper, B-D) or anti-glucagon antibodies (A, lower) and the right images represent the merge signal (Merge, yellow). (A-C) Magnificationx400. (D) Magnificationx1200. Scale bars-50 µm. Representative staining of pancreatic tissues derived from 2–4 animals per each group is shown.
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pone-0074033-g007: Expression of the NKp46 ligand in beta cells derived from P. Obesus.(A)Immunofluorescence staining of pancreatic tissues derived from 8–10 weeks of age, male, P. obesus gerbils fed with LE diet. In the upper images staining for insulin and for NKp46 ligand are shown and in the lower images staining for NKp46 ligand and glucagon are shown. (B) NKp46-Ig and anti-insulin staining of pancreases derived from diabetic (non-fasting blood glucose>250 mg/dl for 2 sequential days) P. obesus gerbils fed with HE diet for 21–24 days. (C) NKp46-Ig and anti-insulin staining of pancreatic tissues derived from diabetic P. obesus gerbils fed with HE diet for 21–24 days followed by 48 hours of LE diet (C, upper) or overnight (O/N) fasting (C, lower). (D) NKp46-Ig and anti-insulin staining of pancreatic tissues derived from non-diabetic P. obesus fed for 21–24 days with HE diet and then injected with Exendin 4 (EX4). For all Figure parts the left images represent staining with NKp46-Ig (NKp46 ligand, green), the middle images represent staining with anti-insulin (A, upper, B-D) or anti-glucagon antibodies (A, lower) and the right images represent the merge signal (Merge, yellow). (A-C) Magnificationx400. (D) Magnificationx1200. Scale bars-50 µm. Representative staining of pancreatic tissues derived from 2–4 animals per each group is shown.

Mentions: P. obesus is a gerbil habituating in African and Mediterranean semi-desert regions [17], [18]. In their native habitat these gerbils feed on low calorie Atriplex halimus plants and are normoglycemic [17], [18], [19]. However, in captivity, under conditions of restrained physical activity and of high energy diet (HE) feeding, P. obesus rapidly develop hyperglycemia, hyperinsulinemia and marked depletion of pancreatic insulin content [17], [18], [19]. To test whether the expression of NKp46 is altered in the nutrition-induced diabetes of P. obesus we first performed co-immunofluorescence staining of pancreatic tissues derived from normoglycemic P. obesus fed a low energy (LE) diet. Consistent with our previous findings in mice and humans [10], [11], the beta cells of P. obesus also expressed the NKp46 ligand, whereas the alpha cells stained negative with NKp46-Ig (Figure 7A). Control staining is shown in Figure S1B.


The expression of the beta cell-derived autoimmune ligand for the killer receptor nkp46 is attenuated in type 2 diabetes.

Gur C, Enk J, Weitman E, Bachar E, Suissa Y, Cohen G, Schyr RB, Sabanay H, Horwitz E, Glaser B, Dor Y, Pribluda A, Hanna JH, Leibowitz G, Mandelboim O - PLoS ONE (2013)

Expression of the NKp46 ligand in beta cells derived from P. Obesus.(A)Immunofluorescence staining of pancreatic tissues derived from 8–10 weeks of age, male, P. obesus gerbils fed with LE diet. In the upper images staining for insulin and for NKp46 ligand are shown and in the lower images staining for NKp46 ligand and glucagon are shown. (B) NKp46-Ig and anti-insulin staining of pancreases derived from diabetic (non-fasting blood glucose>250 mg/dl for 2 sequential days) P. obesus gerbils fed with HE diet for 21–24 days. (C) NKp46-Ig and anti-insulin staining of pancreatic tissues derived from diabetic P. obesus gerbils fed with HE diet for 21–24 days followed by 48 hours of LE diet (C, upper) or overnight (O/N) fasting (C, lower). (D) NKp46-Ig and anti-insulin staining of pancreatic tissues derived from non-diabetic P. obesus fed for 21–24 days with HE diet and then injected with Exendin 4 (EX4). For all Figure parts the left images represent staining with NKp46-Ig (NKp46 ligand, green), the middle images represent staining with anti-insulin (A, upper, B-D) or anti-glucagon antibodies (A, lower) and the right images represent the merge signal (Merge, yellow). (A-C) Magnificationx400. (D) Magnificationx1200. Scale bars-50 µm. Representative staining of pancreatic tissues derived from 2–4 animals per each group is shown.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3757008&req=5

pone-0074033-g007: Expression of the NKp46 ligand in beta cells derived from P. Obesus.(A)Immunofluorescence staining of pancreatic tissues derived from 8–10 weeks of age, male, P. obesus gerbils fed with LE diet. In the upper images staining for insulin and for NKp46 ligand are shown and in the lower images staining for NKp46 ligand and glucagon are shown. (B) NKp46-Ig and anti-insulin staining of pancreases derived from diabetic (non-fasting blood glucose>250 mg/dl for 2 sequential days) P. obesus gerbils fed with HE diet for 21–24 days. (C) NKp46-Ig and anti-insulin staining of pancreatic tissues derived from diabetic P. obesus gerbils fed with HE diet for 21–24 days followed by 48 hours of LE diet (C, upper) or overnight (O/N) fasting (C, lower). (D) NKp46-Ig and anti-insulin staining of pancreatic tissues derived from non-diabetic P. obesus fed for 21–24 days with HE diet and then injected with Exendin 4 (EX4). For all Figure parts the left images represent staining with NKp46-Ig (NKp46 ligand, green), the middle images represent staining with anti-insulin (A, upper, B-D) or anti-glucagon antibodies (A, lower) and the right images represent the merge signal (Merge, yellow). (A-C) Magnificationx400. (D) Magnificationx1200. Scale bars-50 µm. Representative staining of pancreatic tissues derived from 2–4 animals per each group is shown.
Mentions: P. obesus is a gerbil habituating in African and Mediterranean semi-desert regions [17], [18]. In their native habitat these gerbils feed on low calorie Atriplex halimus plants and are normoglycemic [17], [18], [19]. However, in captivity, under conditions of restrained physical activity and of high energy diet (HE) feeding, P. obesus rapidly develop hyperglycemia, hyperinsulinemia and marked depletion of pancreatic insulin content [17], [18], [19]. To test whether the expression of NKp46 is altered in the nutrition-induced diabetes of P. obesus we first performed co-immunofluorescence staining of pancreatic tissues derived from normoglycemic P. obesus fed a low energy (LE) diet. Consistent with our previous findings in mice and humans [10], [11], the beta cells of P. obesus also expressed the NKp46 ligand, whereas the alpha cells stained negative with NKp46-Ig (Figure 7A). Control staining is shown in Figure S1B.

Bottom Line: Following glucose stimulation the NKp46 ligand translocates to the cell membrane and its secretion decreases.We further demonstrate by using several modalities that the unknown NKp46 ligand is not insulin.We demonstrate that the expression of the NKp46 ligand is decreased in all models of T2D studied, suggesting that NKp46 is not involved in T2D.

View Article: PubMed Central - PubMed

Affiliation: The Lautenberg Center for General and Tumor Immunology, The Department of Immunology and Cancer Research, The Institute for Medical Research Israel-Canada, The Hebrew University-Hadassah Medical School, Jerusalem, Israel.

ABSTRACT
NK cells rapidly kill tumor cells, virus infected cells and even self cells. This is mediated via killer receptors, among which NKp46 (NCR1 in mice) is prominent. We have recently demonstrated that in type 1 diabetes (T1D) NK cells accumulate in the diseased pancreas and that they manifest a hyporesponsive phenotype. In addition, we found that NKp46 recognizes an unknown ligand expressed by beta cells derived from humans and mice and that blocking of NKp46 activity prevented diabetes development. Here we investigated the properties of the unknown NKp46 ligand. We show that the NKp46 ligand is mainly located in insulin granules and that it is constitutively secreted. Following glucose stimulation the NKp46 ligand translocates to the cell membrane and its secretion decreases. We further demonstrate by using several modalities that the unknown NKp46 ligand is not insulin. Finally, we studied the expression of the NKp46 ligand in type 2 diabetes (T2D) using 3 different in vivo models and 2 species; mice and gerbils. We demonstrate that the expression of the NKp46 ligand is decreased in all models of T2D studied, suggesting that NKp46 is not involved in T2D.

Show MeSH
Related in: MedlinePlus