1. Volume 124, Issue 3, Pages 725-736 (March 2003)
Macrophage migration inhibitory factor is a critical mediator of severe acute pancreatitis 
Yoshitaka Sakai, Atsushi Masamune, Akihiko Satoh, Jun Nishihira, Tetsuya Yamagiwa, Tooru Shimosegawa 
Gastroenterology 
Volume 124, Issue 3, Pages (March 2003)
DOI: /gast
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2. Fig. 1
Ascitic and serum MIF levels in experimental models of AP. The (A) ascites and (B) serum were collected from the abdominal cavity and the abdominal aorta, respectively, at 0, 1, 2, 3, 6, 9, and 24 hours after the induction of pancreatitis (n = 6 for each time point). MIF levels were determined by ELISA. (A) Ascitic MIF levels during cerulein pancreatitis were 17.1 ± 8.8, 101 ± 12, 104 ± 9.2, 136 ± 15, 120 ± 18, 123 ± 11, 117 ± 15 ng/mL at 0, 1, 2, 3, 6, 9, and 24 hours after the induction, respectively. Ascitic MIF levels during TCA pancreatitis were 17.1 ± 8.8, 265 ± 7, 252 ± 7, 245 ± 10, 196 ± 14, 197 ± 9, 140 ± 9 ng/mL at 0, 1, 2, 3, 6, 9, and 24 hours after the induction, respectively. **P < 0.01 vs. 0 hour. (B) Serum MIF levels during cerulein pancreatitis were 68.6 ± 10, 85 ± 12, 84.3 ± 12, 87.7 ± 22, 119 ± 31, 89.3 ± 6.1, 89.7 ± 24 ng/mL at 0, 1, 2, 3, 6, 9, and 24 hours after induction of AP, respectively. Serum MIF levels during TCA pancreatitis were 68.6 ± 10, ± 13, 168 ± 36, 157 ± 35, 182 ± 23, 200 ± 47, 170 ± 14 ng/mL at 0, 1, 2, 3, 6, 9, and 24 hours after induction of AP, respectively. **P < 0.01 vs. 0 hour.
Gastroenterology  , DOI: ( /gast )
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3. Fig. 2
MIF levels in the organs of normal and TCA pancreatitis rats. MIF levels in the pancreas, the liver, and the lung in the normal rats and in TCA pancreatitis rats (at 6 h after the induction of pancreatitis) were determined by ELISA (n = 6 for each organ). *P < ■, normal; ▨, TCA pancreatitis.
Gastroenterology  , DOI: ( /gast )
Copyright © 2003 American Gastroenterological Association Terms and Conditions

4. Fig. 3
Immunostaining for MIF on the lung of normal and TCA pancreatitis rats. MIF expression in the lung of (A) normal, and (B) TCA pancreatitis rats (at 6 h after the induction) were examined by immunostaining. Original magnification: ×40 objective. Positive immunostaining was observed predominantly within the bronchial epithelial cells in the lung even in normal rats, and the staining was increased in TCA pancreatitis.
Gastroenterology  , DOI: ( /gast )
Copyright © 2003 American Gastroenterological Association Terms and Conditions

5. Fig. 4
Immunostaining for MIF in the peritoneal macrophages from TCA pancreatitis rats. MIF expression in the peritoneal macrophages from TCA pancreatitis was examined by double staining for MIF and ED-1. Adherent cells on the slides from the ascitic fluids were incubated with mouse monoclonal anti-rat mononuclear phagocyte antibody (anti–ED-1) and rabbit polyclonal anti-MIF antibody. Then, samples were incubated with fluorescein isothiocyanate–conjugated sheep anti-mouse IgG and rhodamine red–conjugated goat anti-rabbit IgG. Expression of ED-1 and MIF was evaluated under fluorescent microscopy. Colocalization of MIF and ED-1 expression indicated that peritoneal macrophages were a source of the ascitic MIF. Original magnification: ×40 objective.
Gastroenterology  , DOI: ( /gast )
Copyright © 2003 American Gastroenterological Association Terms and Conditions

6. Fig. 5
Anti-MIF antibody inhibited ascitic fluid–induced IL-8 production in vitro. (A) MIF in the ascitic fluids was neutralized with the anti-rat MIF antibody at the indicated concentrations by incubation at 37°C for 4 hours. Human monocytic THP-1 cells were stimulated with the pretreated ascitic fluids (at the final concentration of 30%) for 24 hours, and IL-8 levels in the culture supernatants were determined by ELISA. *P < 0.05, **P < 0.01 vs. ascitic fluids only. (B) THP-1 cells were treated with recombinant rat and human MIFs at 25 ng/mL for 24 hours. IL-8 levels in the culture supernatants were determined. **P < 0.01 vs. control.
Gastroenterology  , DOI: ( /gast )
Copyright © 2003 American Gastroenterological Association Terms and Conditions

7. Fig. 6
Anti-MIF antibody protected against lethal TCA pancreatitis. Rats were injected intraperitoneally with anti-MIF antibody or nonspecific IgG (control antibody) (A) 1 hour before, (B) immediately after, or (C) 1 hour after the induction of pancreatitis. Anti-MIF antibody given at 1 hour before the induction of pancreatitis significantly improved the survival rate from 44% to 88% at 120 hours after the induction of pancreatitis (P < 0.05, n = 16 for each group). Anti-MIF antibody given immediately after the induction also significantly improved the survival rate from 33% to 92% (P < 0.01, n = 12 for each group). If anti-MIF antibody were given at 1 hour after the induction of pancreatitis, there was a tendency of improved survival rate, but it was not statistically significant (from 39% to 61%, P = 0.32). Treatment with nonspecific rabbit IgG (control antibody), saline, or PBS (vehicle for the antibody) did not affect the survival rates of TCA pancreatitis. *P < 0.05, **P < 0.01 vs. control.
Gastroenterology  , DOI: ( /gast )
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8. Fig. 7
Effects of anti-MIF antibody treatment on the histologic findings in the pancreas. Rats were injected intraperitoneally with anti-MIF antibody 1 hour before the induction of TCA pancreatitis, and the pancreas tissue was obtained 18 hours after the induction of pancreatitis. The gross pathologic appearance of the pancreas was not affected by the antibody treatment (A, control antibody; B, anti-MIF antibody). Original magnification: ×20 objective.
Gastroenterology  , DOI: ( /gast )
Copyright © 2003 American Gastroenterological Association Terms and Conditions

9. Fig. 8
Anti-MIF antibody decreased pulmonary TNF-α levels of TCA pancreatitis rats. Rats were injected intraperitoneally with anti-MIF antibody or nonspecific IgG (control antibody) 1 hour before the induction of TCA pancreatitis. At 18 hours after induction of pancreatitis the lungs were harvested and TNF-α levels were determined by ELISA (n = 6 for each group). Pulmonary TNF-α levels were 48.3 ± 7.5 pg/mg protein in normal rats, 140 ± 15.7 pg/mg protein in the control antibody group, and 71.3 ± 2.3 pg/mg protein in the anti-MIF antibody group. Pulmonary TNF-α levels were significantly increased in TCA pancreatitis, and the increase was attenuated by the treatment with the anti-MIF antibody, but not with control antibody (**P < 0.01). Ab, antibody.
Gastroenterology  , DOI: ( /gast )
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10. Fig. 9
Anti-MIF antibody improved the survival rate of mice with CDE pancreatitis. Female CD-1 mice were fasted for 24 hours, and then received the CDE diet for 48 hours. After a second period of fasting for 24 hours, mice were given normal laboratory chow until the end of the experiment. (A, B) MIF expression in the lung of (A) normal mice and (B) mice with CDE pancreatitis (at 72 h after the onset of CDE diet) were examined by immunostaining. Original magnification: ×40 objective. Positive immunostaining was observed predominantly within the bronchial epithelial cells in the lung even in normal mice, and the staining was increased in CDE pancreatitis. (C) Anti-MIF antibody (at 10 mg/kg) or nonspecific IgG (control antibody) was injected intraperitoneally immediately after the onset of CDE diet. This treatment was repeated every 12 hours. Mice were observed every 6 hours for 7 days. The treatment of anti-MIF antibody significantly improved the survival rate from 16% to 37% (*P < 0.05, n = 43 for each group).
Gastroenterology  , DOI: ( /gast )
Copyright © 2003 American Gastroenterological Association Terms and Conditions

11. Fig. 10
Serum MIF levels were increased in severe AP patients. Serum MIF levels of healthy individuals (control) (n = 12) and of patients with mild AP (n = 28) or severe AP (n = 18) were determined by ELISA. The median serum MIF levels in humans were 26.2 ng/mL (range, 1–70) in mild AP, 45.8 ng/mL in severe AP (range, 20–112), and 18.2 ng/mL (range, 11–34) in healthy controls. The P values for comparison between the groups were for healthy individuals vs. severe AP, for mild AP vs. severe AP, and 0.28 for healthy individuals vs. mild AP. **P < 0.01.
Gastroenterology  , DOI: ( /gast )
Copyright © 2003 American Gastroenterological Association Terms and Conditions