Limits...
Nitric oxide primes pancreatic beta cells for Fas-mediated destruction in insulin-dependent diabetes mellitus.

Stassi G, De Maria R, Trucco G, Rudert W, Testi R, Galluzzo A, Giordano C, Trucco M - J. Exp. Med. (1997)

Bottom Line: Fas is an apoptosis-inducing surface receptor involved in controlling tissue homeostasis and function at multiple sites.Normal human pancreatic beta cells that do not constitutively express Fas, become strongly Fas positive after interleuken (IL)-1beta exposure, and are then susceptible to Fas-mediated apoptosis.NG-monomethyl-L-arginine, an inhibitor of nitric oxide (NO) synthase, prevents IL-1beta-induced Fas expression, whereas the NO donors sodium nitroprusside and nitric oxide releasing compound (NOC)-18, induce functional Fas expression in normal pancreatic beta cells.

View Article: PubMed Central - PubMed

Affiliation: Division of Immunogenetics, Department of Pediatrics, University of Pittsburgh School of Medicine, PA 15213, USA.

ABSTRACT
Fas is an apoptosis-inducing surface receptor involved in controlling tissue homeostasis and function at multiple sites. Here we show that beta cells from the pancreata of newly diagnosed insulin-dependent diabetes mellitus (IDDM) patients express Fas and show extensive apoptosis among those cells located in proximity to Fas ligand-expressing T lymphocytes infiltrating the IDDM islets. Normal human pancreatic beta cells that do not constitutively express Fas, become strongly Fas positive after interleuken (IL)-1beta exposure, and are then susceptible to Fas-mediated apoptosis. NG-monomethyl-L-arginine, an inhibitor of nitric oxide (NO) synthase, prevents IL-1beta-induced Fas expression, whereas the NO donors sodium nitroprusside and nitric oxide releasing compound (NOC)-18, induce functional Fas expression in normal pancreatic beta cells. These findings suggest that NO-mediated upregulation of Fas contributes to pancreatic beta cell damage in IDDM.

Show MeSH

Related in: MedlinePlus

Induction of apoptosis in human pancreatic islet cells by anti-Fas mAb (CH-11). (A) Islet cells incubated for 24 h in tissue culture medium and then exposed for an additional 24 h to 200 ng/ml CH-11 mAb,  or in medium supplemented with IL-1β (50 U/ml) or 0.5 mmol/liter  SNP followed by control IgM, or in medium containing IL-1β or SNP  followed by 10 μg/ml ZB4 or CH-11 mAb, or in medium containing  IL-1β plus 3 mmol/liter L-NMMA followed by CH-11 mAb, were processed for DNA content analysis by staining. Percentages of apoptotic  (i.e., hypodiploid) nuclei are indicated between shift markers, plotted on  log histograms as red fluorescence intensity (x-axis) versus relative nuclei  number (y-axis). (B) Evaluation of cell survival of human α (left) and β  cells (right) cultured in culture medium for 24 h and then for 24 h in medium in the absence of IL-1β, in medium supplemented with IL-1β (50  U/ml; open squares), and control mAb (open circles), or in medium with IL-1β  and CH-11 mAb (closed circles), or in medium with IL-1β plus L-NMMA  and CH-11 mAb (closed triangles). Data shown represent the values at the  indicated time points of a representative experiment out of a total of seven  independent experiments performed.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2199078&req=5

Figure 3: Induction of apoptosis in human pancreatic islet cells by anti-Fas mAb (CH-11). (A) Islet cells incubated for 24 h in tissue culture medium and then exposed for an additional 24 h to 200 ng/ml CH-11 mAb, or in medium supplemented with IL-1β (50 U/ml) or 0.5 mmol/liter SNP followed by control IgM, or in medium containing IL-1β or SNP followed by 10 μg/ml ZB4 or CH-11 mAb, or in medium containing IL-1β plus 3 mmol/liter L-NMMA followed by CH-11 mAb, were processed for DNA content analysis by staining. Percentages of apoptotic (i.e., hypodiploid) nuclei are indicated between shift markers, plotted on log histograms as red fluorescence intensity (x-axis) versus relative nuclei number (y-axis). (B) Evaluation of cell survival of human α (left) and β cells (right) cultured in culture medium for 24 h and then for 24 h in medium in the absence of IL-1β, in medium supplemented with IL-1β (50 U/ml; open squares), and control mAb (open circles), or in medium with IL-1β and CH-11 mAb (closed circles), or in medium with IL-1β plus L-NMMA and CH-11 mAb (closed triangles). Data shown represent the values at the indicated time points of a representative experiment out of a total of seven independent experiments performed.

Mentions: To determine whether Fas is functional and able to induce a death signal in β cells, we incubated IL-1β– or SNP– treated and untreated islet cells with an agonist (CH-11) or a blocking anti-Fas (ZB4) mAb and examined them for evidence of apoptosis. Treatment with CH-11 mAb in the absence of IL-1β or SNP did not trigger apoptosis. By contrast, incubation with CH-11 dramatically increased apoptosis in SNP– or IL-1β–treated-cells (Fig. 3 A). Immunofluorescence staining and fluorescence microscopy analysis revealed that after 24 h, essentially all cells surviving IL-1β and anti-Fas treatment were glucagon-positive α cells, whereas the recovery of insulin-producing cells was extremely low, indicating that only β cells underwent apoptosis (Fig. 3 B). Moreover, L-NMMA was able to completely suppress IL-1β–induced apoptotic cell death, whereas the anti-Fas antagonist ZB4 did not interfere with IL-1β– or SNP–induced apoptosis (Fig. 3 A). Thus, NO mediates both Fas-independent and Fas-dependent β cell apoptosis induced by IL-1β.


Nitric oxide primes pancreatic beta cells for Fas-mediated destruction in insulin-dependent diabetes mellitus.

Stassi G, De Maria R, Trucco G, Rudert W, Testi R, Galluzzo A, Giordano C, Trucco M - J. Exp. Med. (1997)

Induction of apoptosis in human pancreatic islet cells by anti-Fas mAb (CH-11). (A) Islet cells incubated for 24 h in tissue culture medium and then exposed for an additional 24 h to 200 ng/ml CH-11 mAb,  or in medium supplemented with IL-1β (50 U/ml) or 0.5 mmol/liter  SNP followed by control IgM, or in medium containing IL-1β or SNP  followed by 10 μg/ml ZB4 or CH-11 mAb, or in medium containing  IL-1β plus 3 mmol/liter L-NMMA followed by CH-11 mAb, were processed for DNA content analysis by staining. Percentages of apoptotic  (i.e., hypodiploid) nuclei are indicated between shift markers, plotted on  log histograms as red fluorescence intensity (x-axis) versus relative nuclei  number (y-axis). (B) Evaluation of cell survival of human α (left) and β  cells (right) cultured in culture medium for 24 h and then for 24 h in medium in the absence of IL-1β, in medium supplemented with IL-1β (50  U/ml; open squares), and control mAb (open circles), or in medium with IL-1β  and CH-11 mAb (closed circles), or in medium with IL-1β plus L-NMMA  and CH-11 mAb (closed triangles). Data shown represent the values at the  indicated time points of a representative experiment out of a total of seven  independent experiments performed.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2199078&req=5

Figure 3: Induction of apoptosis in human pancreatic islet cells by anti-Fas mAb (CH-11). (A) Islet cells incubated for 24 h in tissue culture medium and then exposed for an additional 24 h to 200 ng/ml CH-11 mAb, or in medium supplemented with IL-1β (50 U/ml) or 0.5 mmol/liter SNP followed by control IgM, or in medium containing IL-1β or SNP followed by 10 μg/ml ZB4 or CH-11 mAb, or in medium containing IL-1β plus 3 mmol/liter L-NMMA followed by CH-11 mAb, were processed for DNA content analysis by staining. Percentages of apoptotic (i.e., hypodiploid) nuclei are indicated between shift markers, plotted on log histograms as red fluorescence intensity (x-axis) versus relative nuclei number (y-axis). (B) Evaluation of cell survival of human α (left) and β cells (right) cultured in culture medium for 24 h and then for 24 h in medium in the absence of IL-1β, in medium supplemented with IL-1β (50 U/ml; open squares), and control mAb (open circles), or in medium with IL-1β and CH-11 mAb (closed circles), or in medium with IL-1β plus L-NMMA and CH-11 mAb (closed triangles). Data shown represent the values at the indicated time points of a representative experiment out of a total of seven independent experiments performed.
Mentions: To determine whether Fas is functional and able to induce a death signal in β cells, we incubated IL-1β– or SNP– treated and untreated islet cells with an agonist (CH-11) or a blocking anti-Fas (ZB4) mAb and examined them for evidence of apoptosis. Treatment with CH-11 mAb in the absence of IL-1β or SNP did not trigger apoptosis. By contrast, incubation with CH-11 dramatically increased apoptosis in SNP– or IL-1β–treated-cells (Fig. 3 A). Immunofluorescence staining and fluorescence microscopy analysis revealed that after 24 h, essentially all cells surviving IL-1β and anti-Fas treatment were glucagon-positive α cells, whereas the recovery of insulin-producing cells was extremely low, indicating that only β cells underwent apoptosis (Fig. 3 B). Moreover, L-NMMA was able to completely suppress IL-1β–induced apoptotic cell death, whereas the anti-Fas antagonist ZB4 did not interfere with IL-1β– or SNP–induced apoptosis (Fig. 3 A). Thus, NO mediates both Fas-independent and Fas-dependent β cell apoptosis induced by IL-1β.

Bottom Line: Fas is an apoptosis-inducing surface receptor involved in controlling tissue homeostasis and function at multiple sites.Normal human pancreatic beta cells that do not constitutively express Fas, become strongly Fas positive after interleuken (IL)-1beta exposure, and are then susceptible to Fas-mediated apoptosis.NG-monomethyl-L-arginine, an inhibitor of nitric oxide (NO) synthase, prevents IL-1beta-induced Fas expression, whereas the NO donors sodium nitroprusside and nitric oxide releasing compound (NOC)-18, induce functional Fas expression in normal pancreatic beta cells.

View Article: PubMed Central - PubMed

Affiliation: Division of Immunogenetics, Department of Pediatrics, University of Pittsburgh School of Medicine, PA 15213, USA.

ABSTRACT
Fas is an apoptosis-inducing surface receptor involved in controlling tissue homeostasis and function at multiple sites. Here we show that beta cells from the pancreata of newly diagnosed insulin-dependent diabetes mellitus (IDDM) patients express Fas and show extensive apoptosis among those cells located in proximity to Fas ligand-expressing T lymphocytes infiltrating the IDDM islets. Normal human pancreatic beta cells that do not constitutively express Fas, become strongly Fas positive after interleuken (IL)-1beta exposure, and are then susceptible to Fas-mediated apoptosis. NG-monomethyl-L-arginine, an inhibitor of nitric oxide (NO) synthase, prevents IL-1beta-induced Fas expression, whereas the NO donors sodium nitroprusside and nitric oxide releasing compound (NOC)-18, induce functional Fas expression in normal pancreatic beta cells. These findings suggest that NO-mediated upregulation of Fas contributes to pancreatic beta cell damage in IDDM.

Show MeSH
Related in: MedlinePlus