Kupffer cells from halothane-exposed guinea pigs carry trifluoroacetylated protein adducts.
The anesthetic, halothane, is bioactivated by the liver cytochrome P450 system to trifluoroacetyl-chloride, which can readily acylate liver protein. Covalent binding of the trifluoroacetyl moiety may result in hapten formation leading to the induction of an immune response and ultimately halothane hepatitis. In this study the presence of trifluoroacetylated-protein adducts in Kupffer cells was investigated to learn how the immune system might come in contact with the proteins. Guinea pigs were exposed to 1.0% halothane, 40% oxygen for 4 h. Kupffer cells were isolated on days 1 through 9 post-exposure, by liver perfusion and purification by elutriation. Using gel electrophoresis and Western blotting techniques, it has been demonstrated that Kupffer cells obtained from halothane-treated guinea pigs, do carry trifluoroacetyl-protein adducts as recognized by an anti-trifluoroacetyl-rabbit serum albumin antibody. Apparent molecular weights of polypeptides bound by trifluoroacetyl were of a wide range, 25-152 kDa. Bands were most prominent in the larger Kupffer cells with more appearing at lower molecular weights. Trifluoroacetyl-protein adducts were not detected in lung, spleen, lymph node or peripheral blood macrophages. This work suggests a role for Kupffer cells in the presentation of altered proteins in the liver to cells of the immune system.
Furst
S M
SM
Department of Anesthesiology, College of Medicine, The University of Arizona, Tucson 85724, USA.
Luedke
D
D
Gandolfi
A J
AJ
D000107:Acetylation; D018685:Anesthetics, Inhalation / Q000008:administration & dosage / Q000633:toxicity*; D000818:Animals; D000917:Antibody Formation; D000936:Antigen-Antibody Complex; D000938:Antigen-Presenting Cells / Q000166:cytology / Q000187:drug effects*; D015153:Blotting, Western; D002469:Cell Separation; D002470:Cell Survival / Q000187:drug effects; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006168:Guinea Pigs; D006221:Halothane / Q000008:administration & dosage / Q000633:toxicity*; D007728:Kupffer Cells / Q000166:cytology / Q000187:drug effects*; D008198:Lymph Nodes / Q000166:cytology / Q000187:drug effects; D008264:Macrophages / Q000166:cytology / Q000187:drug effects; D016676:Macrophages, Alveolar / Q000166:cytology / Q000187:drug effects; D008297:Male; D008970:Molecular Weight; D011817:Rabbits; D012709:Serum Albumin / Q000276:immunology; D013047:Specific Pathogen-Free Organisms; D013154:Spleen / Q000166:cytology / Q000187:drug effects; D014269:Trifluoroacetic Acid / Q000276:immunology / Q000378:metabolism*
D018685:Anesthetics, Inhalation; D000936:Antigen-Antibody Complex; D012709:Serum Albumin; D003577:Cytochrome P-450 Enzyme System; D014269:Trifluoroacetic Acid; D006221:Halothane