Immunohistochemical localization and quantification of the 3-(cystein-S- yl)-acetaminophen protein adduct in acetaminophen hepatotoxicity

• Published in: The American journal of pathology Vol. 138; no. 2; pp. 359 - 371
• Main Authors: Roberts, DW, Bucci, TJ, Benson, RW, Warbritton, AR, McRae, TA, Pumford, NR, Hinson, JA
• Format: Journal Article
• Language: English
• Published: Bethesda, MD ASIP 01.02.1991; American Society for Investigative Pathology
• Subjects: Acetaminophen - analogs & derivatives; Acetaminophen - metabolism; Acetaminophen - toxicity; Animals; Biological and medical sciences; Dose-Response Relationship, Drug; Drug toxicity and drugs side effects treatment; Immunohistochemistry; Liver - drug effects; Liver - metabolism; Liver - pathology; Male; Medical sciences; Mice; Pharmacology. Drug treatments; Time Factors; Tissue Distribution; Immunohistochemistry; Pathology; Toxicity; Animal; Chelating agent; Digestive diseases; Hepatic disease; Technique
• ISSN: 0002-9440

Acetaminophen overdose causes severe hepatotoxicity in humans and laboratory animals, presumably by metabolism to N-acetyl-p-benzoquinone imine: and binding to cysteine groups as 3-(cystein-S-yl)acetaminophen-protein adduct. Antiserum specific for the adduct was used immunohistochemically to demonstrate the formation, distribution, and concentration of this specific adduct in livers of treated mice and was correlated with cell injury as a function of dose and time. Within the liver lobule, immunohistochemically demonstrable adduct occurred in a temporally progressive, central-to-peripheral pattern. There was concordance between immunohistochemical staining and quantification of the adduct in hepatic 10,000g supernate, using a quantitative particle concentration fluorescence immunoassay. Findings include: 1) immunochemically detectable adduct before the appearance of centrilobular necrosis, 2) distinctive lobular zones of adduct localization with subsequent depletion during the progression of toxicity, 3) drug-protein binding in hepatocytes at subhepatotoxic doses and before depletion of total hepatic glutathione, 4) immunohistochemical evidence of drug binding in the nucleus, and 5) adduct in metabolically active and dividing hepatocytes and in macrophagelike cells in the regenerating liver.