title: Neoantigen formation and clastogenic action of HCFC-123 and perchloroethylene in human MCL-5 cells.
abstract: In this study, the metabolic activation of 2,2-dichloro-1,1,1-trifluoroethane (hydrochlorofluorocarbons-123, HCFC-123), halothane or 1,1-dichloro-1-fluoroethane (HCFC-141b) was compared to that of perchloroethylene, using lymphoblastoma derived cell lines expressing human CYP1A1, CYP1A2, CYP2E1, CYP2A6 and CYP3A4 (MCL-5 cells). A dose dependent increase in micronucleus formation was detected over a nominal concentration range of 0.05-2 mM for HCFC-123 and halothane, but this was not seen with HCFC-141b. No dose response for HCFC-123 was seen in a control cHo1 cell line not expressing this cytochrome P450's. Cell lines expressing individual human cytochrome P-450 (CYP) forms were also used to define the enzymes responsible for the clastogenic events and to investigate the formation of immunoreactive protein by microsomal fractions. It was shown that CYP2E1 or CYP2B6 catalysed the clastogenic response, but CYP2D6, CYP3A4, CYP1A2 or CYP1A1 all appeared to be inactive. The formation of neoantigenic trifluoroacetylated protein adducts by microsomal mixtures incubated with HCFC-123 and NADPH was catalysed primarily by CYP2E1 and to a lesser extent by CYP2C19, whereas, only trace levels of immunoreactive protein were seen with microsomes expressing CYP2B6 or CYP2C8. With perchloroethylene as a substrate, the extent of activation was low in comparison with HCFC-123, as judged by the absence of micronuclei formation in the MCL-5 cell line and the weak immunoreactivity of proteins following Western blotting. CYP1A2, CYP2B6 and CYP2C8 appeared to be responsible for perchloroethylene immunoreactivity and in contrast to the findings with the HCFC's, no activation of perchloroethylene by CYP2E1 could be detected. These results show that even though both saturated and unsaturated halocarbons can result in neoantigen formation, there is a marked difference in the specificity of the CYP enzymes involved in their metabolic activation.
authors.affiliation: MRC Toxicology Unit, Hodgkin Building, University of Leicester, Lancaster Road, Leicester LE1 9HN, UK. iw6@le.ac.uk
authors.forename: I N
authors.initials: IN
authors.lastname: White
authors.forename: N
authors.initials: N
authors.lastname: Razvi
authors.forename: A H
authors.initials: AH
authors.lastname: Gibbs
authors.forename: A M
authors.initials: AM
authors.lastname: Davies
authors.forename: M
authors.initials: M
authors.lastname: Manno
authors.forename: C
authors.initials: C
authors.lastname: Zaccaro
authors.forename: F
authors.initials: F
authors.lastname: De Matteis
authors.forename: A
authors.initials: A
authors.lastname: Pähler
authors.forename: W
authors.initials: W
authors.lastname: Dekant
mesh_terms: D000941:Antigens / Q000032:analysis*; D015153:Blotting, Western; D002273:Carcinogens / Q000009:adverse effects*; D017402:Chlorofluorocarbons / Q000009:adverse effects* / Q000276:immunology*; D064227:Chlorofluorocarbons, Ethane; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007945:Leukemia, Lymphoid / Q000473:pathology; D015162:Micronucleus Tests; D013750:Tetrachloroethylene / Q000009:adverse effects* / Q000276:immunology; D014407:Tumor Cells, Cultured
chemical_list: D000941:Antigens; D002273:Carcinogens; D017402:Chlorofluorocarbons; D064227:Chlorofluorocarbons, Ethane; C067411:2,2-dichloro-1,1,1-trifluoroethane; D003577:Cytochrome P-450 Enzyme System; C068320:1,1-dichloro-1-fluoroethane; D013750:Tetrachloroethylene