Human anti-endoplasmic reticulum antibodies in sera of patients with halothane-induced hepatitis are directed against a trifluoroacetylated carboxylesterase.
Previous studies have demonstrated that patients with halothane-induced hepatitis have serum antibodies that are directed against novel liver microsomal neoantigens and have suggested that these neoantigens may play an immunopathological role in development of the patients' liver damage. These investigations have further revealed that the antibodies are directed against distinct polypeptide fractions (100 kDa, 76 kDa, 59 kDa, 57 kDa, 54 kDa) that have been covalently modified by the reactive trifluoroacetyl halide metabolite of halothane. In this paper, the trifluoroacetylated (TFA) 59-kDa neoantigen (59-kDa-TFA) recognized by the patients' antibodies was isolated from liver microsomes of halothane-treated rats by chromatography on an immunoaffinity column of anti-TFA IgG. Antibodies were raised against the 59-kDa-TFA protein and were used to purify the native protein from liver microsomes of untreated rats. Based upon its apparent monomeric molecular mass, NH2-terminal amino acid sequence, catalytic activity, and other physical properties, the protein has been identified as a previously characterized microsomal carboxylesterase (EC 3.1.1.1). A similar strategy may be used to purify and characterized neoantigens associated with other drug toxicities that are believed to have an immunopathological basis.
Satoh
H
H
Laboratory of Chemical Pharmacology, National Heart, Lung and Blood Institute, Bethesda, MD 20892.
Martin
B M
BM
Schulick
A H
AH
Christ
D D
DD
Kenna
J G
JG
Pohl
L R
LR
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D043182:Carboxylesterase; D002265:Carboxylic Ester Hydrolases / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D056486:Chemical and Drug Induced Liver Injury / Q000276:immunology*; D004721:Endoplasmic Reticulum / Q000276:immunology*; D005463:Fluoroacetates / Q000378:metabolism*; D006221:Halothane / Q000494:pharmacology; D006801:Humans; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D008970:Molecular Weight; D011485:Protein Binding; D051381:Rats; D011919:Rats, Inbred Strains; D014269:Trifluoroacetic Acid / Q000378:metabolism* / Q000494:pharmacology
D000906:Antibodies; D005463:Fluoroacetates; D014269:Trifluoroacetic Acid; D002265:Carboxylic Ester Hydrolases; D043182:Carboxylesterase; D006221:Halothane