title: Monoclonal antibodies against toluene diisocyanate haptenated proteins from vapor-exposed mice.
abstract: Toluene diisocyanate (TDI) is an industrially important polymer cross-linker used in the production of polyurethane. Workplace exposure to TDI and other diisocyanates is reported to be a leading cause of low molecular weight-induced occupational asthma (OA). Currently we have a limited understanding of the pathogenesis of OA. Monoclonal antibodies (MAbs) that recognize TDI bound proteins would be valuable tools/reagents, both in exposure monitoring and in TDI-induced asthma research. We sought to develop toluene diisocyanate (TDI)-specific MAbs for potential use in the development of standardized immunoassays for exposure and biomarker assessments. Mice were exposed 4 h/day for 12 consecutive weekdays to 50 ppb, 2,4;2,6 TDI vapor (80/20 mixture). Splenocytes were isolated 24 h after the last exposure for hybridoma production. Hybridomas were screened in a solid-phase indirect enzyme-linked immunosorbent assay (ELISA) against a 2,4 TDI-human serum albumin (2,4 TDI-HSA) protein conjugate. Three hybridomas producing 2,4 TDI-HSA reactive IgM MAbs were obtained. The properties of these MAbs (isotype and reactivity to various protein-isocyanate conjugate epitopes) were characterized using ELISA, dot blot, and Western blot analyses. Western blot analyses demonstrated that some TDI conjugates form inter- and intra-molecular links, resulting in multimers and a change in the electrophoretic mobility of the conjugate. These antibodies may be useful tools for the isolation of endogenous diisocyanate-modified proteins after natural or experimental exposures and for characterization of the toxicity of specific dNCOs.
authors.affiliation: Allergy and Clinical Immunology, National Institute for Occupational Safety and Health, Morgantown, West Virginia 26505-2888, USA.
authors.forename: Tinashe B
authors.initials: TB
authors.lastname: Ruwona
authors.forename: Victor J
authors.initials: VJ
authors.lastname: Johnson
authors.forename: Detlef
authors.initials: D
authors.lastname: Schmechel
authors.forename: Reuben H
authors.initials: RH
authors.lastname: Simoyi
authors.forename: Donald
authors.initials: D
authors.lastname: Beezhold
authors.forename: Paul D
authors.initials: PD
authors.lastname: Siegel
mesh_terms: D000280:Administration, Inhalation; D000818:Animals; D000911:Antibodies, Monoclonal / Q000145:classification / Q000276:immunology*; D000918:Antibody Specificity; D001249:Asthma / Q000139:chemically induced / Q000175:diagnosis; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006241:Haptens / Q000276:immunology*; D006801:Humans; D006825:Hybridomas; D015151:Immunoblotting; D007075:Immunoglobulin M; D017953:Isocyanates / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D012709:Serum Albumin; D014051:Toluene 2,4-Diisocyanate / Q000008:administration & dosage / Q000276:immunology* / Q000633:toxicity
chemical_list: D000911:Antibodies, Monoclonal; D006241:Haptens; D007075:Immunoglobulin M; D017953:Isocyanates; D012709:Serum Albumin; D014051:Toluene 2,4-Diisocyanate