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Protective effect of monoclonal antibodies to adhesion molecules on rat liver ischemia–reperfusion injury
Protective Effect of Monoclonal Antibodies to Adhesion Molecules on Rat Liver Ischemia–Reperfusion Injury S. Marubayashi, Y. Oshiro, A. Fukuma, K. Okada, T. Maeda, and K. Dohi
T
HE SOURCE of reactive oxygen species during ischemia or subsequent reperfusion remains controversial. Polymorphonuclear neutrophils (PMN) are assumed to be a major source of active oygen species that cause oxidative injury associated with hepatic ischemia and reperfusion. We have shown that cellular injury in hepatic ischemia followed by reperfusion can be explained by free radical reaction processes and especially reperfusion.1 The present study was undertaken to determine whether PMN can contribute to hepatic ischemia-reperfusion injury, and pretreatment with monoclonal antibodies (MAbs) to intracellular adhesion molecule—1 (ICAM-1) (IA-29), leukocyte function associated antigen-1 (LFA-1) (WT-1) and CD-18 (WT-3) could improve energy metabolism and prolong the viability of the organ. MATERIALS AND METHODS Male Wistar rats were used. Rat liver ischemia was induced by clamping blood vessels supplying median and left lateral hepatic lobes. After reperfusion, the infiltration of PMN in the liver was counted histologically and the expression of ICAM-1 was examined immunohistologically. IA-29 plus WT-1 or IA-29 plus WT-3 at a dose of 0.5 mg/kg of body weight were injected intravenously 5 minutes before inducing ischemia. To determine the effect of MAbs on the survival rate, total hepatic ischemia was induced by clamping the hepatic artery, portal vein, and bile duct after making porta-femoral shunt.1
RESULTS
Although ischemia of the liver for 90 minutes did not permit survival of the animals, pretreatment with IA-29 plus WT-1 increased the survival rate to 57% (four of seven). Pretreatment with IA-29 failed to increase the survival rate (2/7). The number of PMN in the liver increased continuously up to 24 hours after reperfusion following 90-minute ischemia, and the expression of ICAM-1 was enhanced 4 hours after reperfusion. This is accompanied by a low recovery of hepatic adenosine triphosphate (ATP) and, on
0041-1345/99/$–see front matter PII S0041-1345(98)01901-0
the contrary, a marked increase in lipid peroxide in the reperfused liver. Pretreatment with MAbs suppressed the infiltration of PMN and the elevation of lipid peroxide, and enhanced the recovery of hepatic ATP 6, 12, and 24 hours after reperfusion. Pretreatment with MAbs also prevented the rise in serum alanine transaminase (ALT) after reperfusion. DISCUSSION
Evidence presented in this article suggests that PMN contribute to ischemia-reperfusion injury in the liver 4 hours and more later period after reperfusion. Jaeschke et al2 reported that Kupffer’s cells, and PMN at a later stage seem to be mainly responsible for the post-ischmeic oxidant stress. Some investigators reported that intraportal injection of ICAM-1 was effective in the prevention of hepatic ischemia-reperfusion injury, but contradictory result has also been reported.3 Therefore, further study are necessary to determine optimal dosage, administration time, injection route, and combination of MABs to obtain the most effective protection with this treatment. In our study MAbs to IA-29 and WT-1 or IA-29 and WT-3 appear to be a promising treatment for ischmeia-reperfusion injury of the liver. REFERENCES 1. Marubayashi S, Dohi K, Sugino K, et al: Ann NY Acad Sci 570:208, 1989 2. Jaeschke H, Farhood A, Bautista AP, et al: Hepatology 17:915, 1993 3. Omura T, Ishikura H, Nakajima Y, et al: Transplant Proc 25:2904, 1993 From the Department of Surgery, Hiroshima University School of Medicine, Hiroshima, Japan. Address reprint requests to Dr Marubayashi, Department of Surgery, Hiroshima University School of Medicine, 1-2-3 Kasumi, Hiroshima 734-8551, Japan.
© 1999 by Elsevier Science Inc. 655 Avenue of the Americas, New York, NY 10010
1054
Transplantation Proceedings, 31, 1054 (1999)
T
HE SOURCE of reactive oxygen species during ischemia or subsequent reperfusion remains controversial. Polymorphonuclear neutrophils (PMN) are assumed to be a major source of active oygen species that cause oxidative injury associated with hepatic ischemia and reperfusion. We have shown that cellular injury in hepatic ischemia followed by reperfusion can be explained by free radical reaction processes and especially reperfusion.1 The present study was undertaken to determine whether PMN can contribute to hepatic ischemia-reperfusion injury, and pretreatment with monoclonal antibodies (MAbs) to intracellular adhesion molecule—1 (ICAM-1) (IA-29), leukocyte function associated antigen-1 (LFA-1) (WT-1) and CD-18 (WT-3) could improve energy metabolism and prolong the viability of the organ. MATERIALS AND METHODS Male Wistar rats were used. Rat liver ischemia was induced by clamping blood vessels supplying median and left lateral hepatic lobes. After reperfusion, the infiltration of PMN in the liver was counted histologically and the expression of ICAM-1 was examined immunohistologically. IA-29 plus WT-1 or IA-29 plus WT-3 at a dose of 0.5 mg/kg of body weight were injected intravenously 5 minutes before inducing ischemia. To determine the effect of MAbs on the survival rate, total hepatic ischemia was induced by clamping the hepatic artery, portal vein, and bile duct after making porta-femoral shunt.1
RESULTS
Although ischemia of the liver for 90 minutes did not permit survival of the animals, pretreatment with IA-29 plus WT-1 increased the survival rate to 57% (four of seven). Pretreatment with IA-29 failed to increase the survival rate (2/7). The number of PMN in the liver increased continuously up to 24 hours after reperfusion following 90-minute ischemia, and the expression of ICAM-1 was enhanced 4 hours after reperfusion. This is accompanied by a low recovery of hepatic adenosine triphosphate (ATP) and, on
0041-1345/99/$–see front matter PII S0041-1345(98)01901-0
the contrary, a marked increase in lipid peroxide in the reperfused liver. Pretreatment with MAbs suppressed the infiltration of PMN and the elevation of lipid peroxide, and enhanced the recovery of hepatic ATP 6, 12, and 24 hours after reperfusion. Pretreatment with MAbs also prevented the rise in serum alanine transaminase (ALT) after reperfusion. DISCUSSION
Evidence presented in this article suggests that PMN contribute to ischemia-reperfusion injury in the liver 4 hours and more later period after reperfusion. Jaeschke et al2 reported that Kupffer’s cells, and PMN at a later stage seem to be mainly responsible for the post-ischmeic oxidant stress. Some investigators reported that intraportal injection of ICAM-1 was effective in the prevention of hepatic ischemia-reperfusion injury, but contradictory result has also been reported.3 Therefore, further study are necessary to determine optimal dosage, administration time, injection route, and combination of MABs to obtain the most effective protection with this treatment. In our study MAbs to IA-29 and WT-1 or IA-29 and WT-3 appear to be a promising treatment for ischmeia-reperfusion injury of the liver. REFERENCES 1. Marubayashi S, Dohi K, Sugino K, et al: Ann NY Acad Sci 570:208, 1989 2. Jaeschke H, Farhood A, Bautista AP, et al: Hepatology 17:915, 1993 3. Omura T, Ishikura H, Nakajima Y, et al: Transplant Proc 25:2904, 1993 From the Department of Surgery, Hiroshima University School of Medicine, Hiroshima, Japan. Address reprint requests to Dr Marubayashi, Department of Surgery, Hiroshima University School of Medicine, 1-2-3 Kasumi, Hiroshima 734-8551, Japan.
© 1999 by Elsevier Science Inc. 655 Avenue of the Americas, New York, NY 10010
1054
Transplantation Proceedings, 31, 1054 (1999)