Immunochemical detection of protein adducts in mice treated with trichloroethylene.
Trichloroethylene has been shown to produce tumors in rodents and is a suspect human carcinogen. In addition, a number of case reports raise the possibility that trichloroethylene can induce an autoimmune disorder known as systemic sclerosis. To investigate whether covalent binding of reactive trichloroethylene metabolites may be involved in the mechanisms underlying these toxic responses, we have developed a polyclonal antibody that can recognize trichloroethylene--protein adducts in tissues. The antibody was prepared by immunizing a rabbit with dichloroacetic anhydride-modified keyhole limpet hemocyanin. Enzyme-linked immunosorbent assay data indicated that the serum antibody recognized dichloroacetic anhydride-modified rabbit serum albumin, but not unmodified protein. In addition, N epsilon-dichloroacetyl-L-lysine was the most potent inhibitor of antibody binding to dichloroacetic anhydride-modified rabbit serum albumin, indicating that the antibody recognizes primarily dichloroacetylated lysine residues. Immunoblots revealed the presence of two major trichloroethylene adducts at 50 and 100 kDa in liver microsomal fractions from male B6C3/F1 mice treated with trichloroethylene. The formation of trichloroethylene adducts was both dose and time dependent. Furthermore, the 50-kDa adduct was found to comigrate on a polyacrylamide gel with cytochrome P450 2E1. These data show that reactive metabolites of trichloroethylene are formed in vivo and bind covalently to discrete proteins in mouse liver. The data also suggest that one of the protein targets is cytochrome P450 2E1. Further studies will be necessary to elucidate the relationship between covalent binding of trichloroethylene and trichloroethylene toxicity.
Halmes
N C
NC
Division of Toxicology, University of Arkansas for Medical Sciences, Little Rock 72205-7199, USA.
McMillan
D C
DC
Oatis
J E
JE
Pumford
N R
NR
D000818:Animals; D000906:Antibodies / Q000737:chemistry; D000918:Antibody Specificity; D019392:Cytochrome P-450 CYP2E1 / Q000737:chemistry / Q000276:immunology; D003999:Dichloroacetic Acid / Q000276:immunology; D015151:Immunoblotting; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008862:Microsomes, Liver / Q000276:immunology; D011485:Protein Binding; D012709:Serum Albumin / Q000276:immunology / Q000378:metabolism*; D014241:Trichloroethylene / Q000008:administration & dosage / Q000737:chemistry* / Q000276:immunology
D000906:Antibodies; D012709:Serum Albumin; D014241:Trichloroethylene; D003999:Dichloroacetic Acid; D019392:Cytochrome P-450 CYP2E1