1. Volume 133, Issue 1, Pages 195-206 (July 2007)
Sera of Patients With Celiac Disease and Neurologic Disorders Evoke a Mitochondrial- Dependent Apoptosis In Vitro 
Elisabetta Cervio, Umberto Volta, Manuela Verri, Federica Boschi, Ornella Pastoris, Alessandro Granito, Giovanni Barbara, Claudia Parisi, Cristina Felicani, Marcello Tonini, Roberto De Giorgio 
Gastroenterology 
Volume 133, Issue 1, Pages (July 2007)
DOI: /j.gastro
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2. Figure 1
Neuronal apoptosis induced by antineuronal antibody–positive sera of patients with celiac disease and neurologic impairment (celiac disease with neurologic disorder and antineuronal antibodies, CD/ND/NA) in SH-Sy5Y cells. (A) Representative microphotographs showing TUNEL (green fluorochrome) (arrows). Calibration bar = 15 μm. (B) Quantitative analysis of neuroblastoma cells exposed to antineuronal antibody–positive sera showing a significantly greater percentage of TUNEL-positive nuclei compared with antineuronal antibody–negative celiac disease sera (celiac disease without neurologic disorder and without antineuronal antibodies, CD/WND/WNA), control sera (blood donors), and FCS (n = 4 experiments). Data are expressed as mean ± SEM (n = 6–10 subjects); *P < .05, **P < .001.
Gastroenterology  , DOI: ( /j.gastro )
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3. Figure 2
Time course analysis showing a significantly greater number of apoptotic SH-Sy5Y cells evoked by antineuronal antibody–positive celiac disease sera (celiac disease with neurologic disorder and antineuronal antibodies, CD/ND/NA) at 12 and 24 hours, but not at 6 hours, as compared with antineuronal antibody–negative celiac disease sera (celiac disease without neurologic disorder and without antineuronal antibodies, CD/WND/WNA), control sera, and FCS (n = 4 experiments). Data are expressed as mean ± SEM (n = 6–10 subjects); *P < .05, **P < .001.
Gastroenterology  , DOI: ( /j.gastro )
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4. Figure 3
(Top panel) A representative example of caspase-3 intense cytoplasmic staining in cells exposed to antineuronal antibody–positive celiac disease sera (CD/ND/NA) compared with controls. Quantitative data show a significantly greater number of caspase-3–positive neuroblastoma cells exposed to antineuronal antibody–positive celiac disease sera compared with antineuronal antibody–negative celiac disease sera (CD/WND/WNA), control sera, and FCS (n = 4 experiments). (Bottom panel) Similar results with apaf-1 immunolabeling and related quantitative results. Data are expressed as mean ± SEM (n = 6–10 subjects); **P < Calibration bars = 10 μm.
Gastroenterology  , DOI: ( /j.gastro )
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5. Figure 4
Western blot of total proteins of SH-Sy5Y cells showing (A) caspase-9 but not (B) caspase-8 activation after 12 hours of exposure to FCS (n = 4 experiments), tumor necrosis factor–related apoptosis-inducing ligand (Trail), control sera, antineuronal antibody–positive (CD/ND/NA), and antineuronal antibody–negative (CD/WND/WNA) celiac disease sera.
Gastroenterology  , DOI: ( /j.gastro )
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6. Figure 5
The specific activity of citrate synthase (CS) after 6 or 12 hours of exposure to antineuronal antibody–positive celiac disease sera (CD/ND/NA) was significantly higher than controls. In contrast, the specific activity of NADH-ubiquinone oxidoreductase after 12 hours of exposure to antineuronal antibody–positive celiac disease sera (CD/ND/NA) was significantly lower than controls. No significant differences in specific enzymatic activities of succinate dehydrogenase (SDH) and cytochrome oxidase (COX) were observed after 6 and 12 hours of exposure to either FCS or antineuronal antibody–positive celiac disease sera (CD/ND/NA).
Gastroenterology  , DOI: ( /j.gastro )
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7. Figure 6
Cytochrome c release from mitochondria induced by exposure of SH-Sy5Y cells to antineuronal antibody–positive celiac disease sera (CD/ND/NA). (A) Cells treated with control sera showing cytochrome c immunoreactive dot-like structures. (B) Cells treated with antineuronal antibody–positive celiac disease sera (CD/ND/NA) for 16 hours showed a diffuse cytoplasmic cytochrome c localization as exemplified in the merge. Calibration bar for A and B = 16 μm. (C) Western blot of cytosolic protein (CP) and membrane protein (MP) fractions of SH-Sy5Y cells showed a clear cytochrome c release into the cytosol after 16 hours of exposure to antineuronal antibody–positive celiac disease sera (CD/ND/NA), as compared with FCS, control sera, and antineuronal antibody–negative celiac disease sera (CD/WND/WNA). The absence of cytochrome oxidase IV (COX IV) from cytosolic fraction excludes a mitochondrial contamination.
Gastroenterology  , DOI: ( /j.gastro )
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8. Figure 7
Translocation of Bax from cytosol to mitochondria after 16 hours of exposure of neuroblastoma cells to control sera (sequence of pictures in A) or antineuronal antibody–positive celiac disease sera (CD/ND/NA) (sequence of pictures in B). Arrows in pictures in B indicate Bax translocation from cytosol to mitochondria as exemplified in the merge. Calibration bar for A = 16 μm; calibration bar for B = 20 μm. (C) Western blot of cytosolic protein (CP) and membrane protein (MP) fractions of SH-Sy5Y cells showed Bax translocation from cytosol to mitochondria after 16 hours of exposure to antineuronal antibody–positive celiac disease sera (CD/ND/NA) compared with FCS, control sera, and antineuronal antibody–negative celiac disease sera (CD/WND/WNA). The absence of cytochrome oxidase IV (COX IV) from cytosolic fraction excludes a mitochondrial contamination.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2007 AGA Institute Terms and Conditions