Treatment with neutralising antibody against cytokine induced neutrophil chemoattractant (CINC) protects rats against acute pancreatitis associated lung injury

Abstract

Background: Lung injury manifest clinically as adult respiratory distress syndrome (ARDS) is a common cause of morbidity and mortality following acute pancreatitis (AP). Neutrophils play a critical role in the progression of AP to ARDS. C-x-C chemokines are potent neutrophil chemoattractants and activators and have been implicated in AP.

Aims: To evaluate the effect of blocking the C-x-C chemokine, cytokine induced neutrophil chemoattractant (CINC), in AP on pancreatic inflammation and the associated lung injury in rats.

Methods: AP was induced by hourly intraperitoneal injections of caerulein. Goat anti-CINC antibody was administered either before or after starting caerulein injections to evaluate the prophylactic and therapeutic effects, respectively. Severity of AP was determined by measuring plasma amylase, pancreatic water content, and pancreatic myeloperoxidase (MPO) activity as a measure of neutrophil sequestration in the pancreas. Lung injury was determined by measurement of pulmonary microvascular permeability and lung MPO activity.

Results: Treatment with anti-CINC antibody had little effect on caerulein induced pancreatic damage. However, it reduced the caerulein mediated increase in lung MPO activity as well as lung microvascular permeability when administered either prophylactically (lung MPO (fold increase over control): 1.53 (0.21) v. 3.30 (0.46), p<0.05; microvascular permeability (L/P%): 0.42 (0.07) v. 0.77 (0.11), p<0.05) or therapeutically (lung MPO (fold increase over control): 2.13 (0.10) v 4.42 (0.65), p<0.05; microvascular permeability (L/P%): 0.31 (0.05) v 0.79 (0.13), p<0.05).

Conclusion: Treatment with anti-CINC antibody afforded significant protection against pancreatitis associated lung injury. These results suggest that CINC plays an important role in the systemic inflammatory response in AP.

Figure 1

Effects of prophylactic treatment with anti-cytokine induced neutrophil chemoattractant antibody (anti-CINC antibody) on acute pancreatitis. The results shown are mean (SEM) for eight animals in each group. The results are as follows: plasma amylase (U/l)—control 1593 (54); placebo (Pbo) followed by caerulein (Caer) 20 206 (2374); anti-CINC antibody (Ab) followed by Caer 24 266 (1391); pancreatic water content (% of wet weight)—control 69.7 (1.2); Pbo followed by Caer 86.4 (1.0); Ab followed by Caer 84.1 (1.4); pancreatic myeloperoxidase (MPO) activity (fold increase over control)—control 1.0 (0.26); Pbo followed by Caer 9.72 (1.43); Ab followed by Caer 3.67 (0.38). *p<0.05, antibody treated animals with acute pancreatitis compared with placebo group.

Figure 2

Effects of therapeutic treatment with anti-cytokine induced neutrophil chemoattractant antibody (anti-CINC antibody) on acute pancreatitis. The results shown are the mean (SEM) for eight animals in each group. The results are as follows: plasma amylase (U/l)—control 1593 (54); caerulein (Caer) followed by placebo (Pbo) 23 860 (3377); Caer followed by anti-CINC antibody (Ab) 25 046 (5337); pancreatic water content (% of wet weight)—control 69.7 (1.2); Caer followed by Pbo 86.1 (0.5); Caer followed by Ab 83.8 (0.9); pancreatic myeloperoxidase (MPO) activity (fold increase over control)—control 1.0 (0.26); Caer followed by Pbo 8.86 (1.8); Caer followed by Ab 2.38 (0.3). *p<0.05, antibody treated animals with acute pancreatitis compared with placebo group.

Figure 3

Morphological changes in rat pancreas on induction of acute pancreatitis with/without treatment with anti-cytokine induced neutrophil chemoattractant antibody (anti-CINC antibody). (A) Control: no pancreatitis. (B) Caerulein induced acute pancreatitis with placebo. (C) Caerulein induced acute pancreatitis with prophylactic treatment with anti-CINC antibody. (D) Caerulein induced acute pancreatitis with therapeutic treatment with anti-CINC antibody.

Figure 4

Effects of prophylactic treatment with anti-cytokine induced neutrophil chemoattractant antibody (anti-CINC antibody) on acute pancreatitis associated lung injury. The results shown are mean (SEM) for eight animals in each group. The results are as follows: pulmonary microvascular permeability (L/P%)—control 0.15 (0.02); placebo (Pbo) followed by caerulein (Caer) 0.78 (0.11); anti-CINC antibody (Ab) followed by Caer 0.42 (0.04); lung myeloperoxidase (MPO) activity (fold increase over control)—control 1.0 (0.26); Pbo followed by Caer 3.30 (0.46); Ab followed by Caer 1.53 (0.21). *p<0.05, antibody treated animals with acute pancreatitis compared with placebo group group. L/P, lavage to plasma FITC-albumin fluorescence ratio.

Figure 5

Effects of therapeutic treatment with anti-cytokine induced neutrophil chemoattractant antibody (anti-CINC antibody) on acute pancreatitis associated lung injury. The results shown are mean (SEM) for eight animals in each group. The results are as follows: pulmonary microvascular permeability (L/P%)—control 0.15 (0.02); caerulein (Caer) followed by placebo (Pbo) 0.79 (0.12); Caer followed by anti-CINC antibody (Ab) 0.31 (0.05); lung myeloperoxidase (MPO) activity (fold increase over control)—control 1.0 (0.26); Caer followed by Pbo 4.42 (0.64); Caer followed by Ab 2.13 (0.10). *p<0.05, antibody treated animals with acute pancreatitis compared with placebo group. L/P, lavage to plasma FITC-albumin fluorescence ratio.

Figure 6

Morphological changes in rat lung on induction of acute pancreatitis with/without treatment with anti-cytokine induced neutrophil chemoattractant antibody (anti-CINC antibody). (A) Control: no pancreatitis. (B) Caerulein induced acute pancreatitis with placebo. (C) Caerulein induced acute pancreatitis with prophylactic treatment with anti-CINC antibody. (D) Caerulein induced acute pancreatitis with therapeutic treatment with anti-CINC antibody.