Chlorotyrosine protein adducts are reliable biomarkers of neutrophil-induced cytotoxicity in vivo

Persistent Link:
http://hdl.handle.net/10150/610122
Title:
Chlorotyrosine protein adducts are reliable biomarkers of neutrophil-induced cytotoxicity in vivo
Author:
Gujral, Jaspreet; Hinson, Jack; Jaeschke, Hartmut
Affiliation:
Department of Pharmacology and Toxicology, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205, USA; Liver Research Institute, University of Arizona, Tucson, Arizona 85724, USA
Issue Date:
2004
Publisher:
BioMed Central
Citation:
Comparative Hepatology 2004, 3(Suppl 1):S48 http://www.comparative-hepatology.com/content/3/S1/S48
Journal:
Comparative Hepatology
Rights:
©  Gujral et al; licensee BioMed Central Ltd 2004
Collection Information:
This item is part of the UA Faculty Publications collection. For more information this item or other items in the UA Campus Repository, contact the University of Arizona Libraries at repository@u.library.arizona.edu.
Abstract:
INTRODUCTION:A limitation for investigating the pathophysiological role of neutrophils in vivo is the lack of a reliable biomarker for neutrophil cytotoxicity in the liver. Therefore, we investigated if immunohistochemical detection of chlorotyrosine protein adducts can be used as a specific footprint for generation of neutrophil-derived hypochlorous acid in vivo.METHODS:C3Heb/FeJ mice were treated with 100 micrograms/kg endotoxin (ET) alone or in combination with 700 mg/kg galactosamine (Gal/ET). Some animals received additionally two doses of 10 mg/kg of the pancaspase inhibitor Z-VAD-fmk. An antibody against chlorotyrosine was used for the immunohistochemical analysis.RESULTS:At 6 h after Gal/ET, hepatocellular apoptosis was evident without increase in plasma ALT activities. Neutrophils accumulated in sinusoids but there was no evidence for chlorotyrosine staining. At 7 h after Gal/ET, about 54% of the sequestered neutrophils had extravasated, there was extensive necrosis and increased plasma ALT activities. Extensive immunostaining for chlorotyrosine, mainly colocalized with neutrophils, could be observed. Treatment with Z-VAD-fmk eliminated apoptosis, necrosis and the increase in plasma ALT values. Neutrophil extravasation was prevented but the overall number of neutrophils in the liver was unchanged. Chlorotyrosine staining was absent in these samples. After ET alone (7 h), sinusoidal neutrophil accumulation was similar to Gal/ET treatment but there was no apoptosis, neutrophil extravasation, ALT release or chlorotyrosine staining.CONCLUSIONS:Chlorotyrosine staining in liver samples correlated well with evidence of neutrophil-induced liver injury in the endotoxemia model. These results indicate that assessment of chlorotyrosine protein adduct formation by immunohistochemistry could be a useful marker of neutrophil-induced liver cell injury in vivo.
EISSN:
1476-5926
DOI:
10.1186/1476-5926-2-S1-S48
Version:
Final published version
Additional Links:
http://www.comparative-hepatology.com/content/3/S1/S48

Full metadata record

DC FieldValue Language
dc.contributor.authorGujral, Jaspreeten
dc.contributor.authorHinson, Jacken
dc.contributor.authorJaeschke, Hartmuten
dc.date.accessioned2016-05-20T08:59:05Z-
dc.date.available2016-05-20T08:59:05Z-
dc.date.issued2004en
dc.identifier.citationComparative Hepatology 2004, 3(Suppl 1):S48 http://www.comparative-hepatology.com/content/3/S1/S48en
dc.identifier.doi10.1186/1476-5926-2-S1-S48en
dc.identifier.urihttp://hdl.handle.net/10150/610122-
dc.description.abstractINTRODUCTION:A limitation for investigating the pathophysiological role of neutrophils in vivo is the lack of a reliable biomarker for neutrophil cytotoxicity in the liver. Therefore, we investigated if immunohistochemical detection of chlorotyrosine protein adducts can be used as a specific footprint for generation of neutrophil-derived hypochlorous acid in vivo.METHODS:C3Heb/FeJ mice were treated with 100 micrograms/kg endotoxin (ET) alone or in combination with 700 mg/kg galactosamine (Gal/ET). Some animals received additionally two doses of 10 mg/kg of the pancaspase inhibitor Z-VAD-fmk. An antibody against chlorotyrosine was used for the immunohistochemical analysis.RESULTS:At 6 h after Gal/ET, hepatocellular apoptosis was evident without increase in plasma ALT activities. Neutrophils accumulated in sinusoids but there was no evidence for chlorotyrosine staining. At 7 h after Gal/ET, about 54% of the sequestered neutrophils had extravasated, there was extensive necrosis and increased plasma ALT activities. Extensive immunostaining for chlorotyrosine, mainly colocalized with neutrophils, could be observed. Treatment with Z-VAD-fmk eliminated apoptosis, necrosis and the increase in plasma ALT values. Neutrophil extravasation was prevented but the overall number of neutrophils in the liver was unchanged. Chlorotyrosine staining was absent in these samples. After ET alone (7 h), sinusoidal neutrophil accumulation was similar to Gal/ET treatment but there was no apoptosis, neutrophil extravasation, ALT release or chlorotyrosine staining.CONCLUSIONS:Chlorotyrosine staining in liver samples correlated well with evidence of neutrophil-induced liver injury in the endotoxemia model. These results indicate that assessment of chlorotyrosine protein adduct formation by immunohistochemistry could be a useful marker of neutrophil-induced liver cell injury in vivo.en
dc.language.isoenen
dc.publisherBioMed Centralen
dc.relation.urlhttp://www.comparative-hepatology.com/content/3/S1/S48en
dc.rights©  Gujral et al; licensee BioMed Central Ltd 2004en
dc.titleChlorotyrosine protein adducts are reliable biomarkers of neutrophil-induced cytotoxicity in vivoen
dc.typeArticleen
dc.identifier.eissn1476-5926en
dc.contributor.departmentDepartment of Pharmacology and Toxicology, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205, USAen
dc.contributor.departmentLiver Research Institute, University of Arizona, Tucson, Arizona 85724, USAen
dc.identifier.journalComparative Hepatologyen
dc.description.collectioninformationThis item is part of the UA Faculty Publications collection. For more information this item or other items in the UA Campus Repository, contact the University of Arizona Libraries at repository@u.library.arizona.edu.en
dc.eprint.versionFinal published versionen
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