Monoclonal Antibodies Against Toluene Diisocyanate Haptenated Proteins from Vapor-Exposed Mice

Details

• Personal Author:
  Beezhold, Donald H. ; Johnson VJ ; Ruwona TB ; Schmechel D ; Siegel, Paul D. ; Simoyi RH
• Description:
  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. [Description provided by NIOSH]
• Subjects:
  Antibodies Antigens Biochemistry Biological Factors Environmental Monitoring Hazardous Substances Hypersensitivity Immune System Laboratories Lung Lung Diseases Manufacturing And Industrial Facilities Occupational Exposure Occupational Health Respiratory System Respiratory Tract Diseases Safety

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• Keywords:
  Antibody-response Biochemical-analysis Biological-effects Biological-factors Biological-systems Breathing Chemical-hypersensitivity Chemical-industry-workers Exposure-assessment Exposure-levels Exposure-methods Immune-reaction Immunochemistry Laboratory-animals Laboratory-testing Occupational-exposure Occupational-health Occupational-respiratory-disease Pulmonary-disorders Pulmonary-system-disorders Toxic-effects

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• ISSN:
  1554-0014
• Document Type:
  Text
• Genre:
  Journal Article
• Place as Subject:
  Oregon ; OSHA Region 10 ; OSHA Region 3 ; West Virginia
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Division:
  HELD - Health Effects Laboratory Division
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Pages in Document:
  221-229
• Volume:
  29
• Issue:
  3
• NIOSHTIC Number:
  nn:20037054
• Citation:
  Hybridoma 2010 Jun; 29(3):221-229
• Contact Point Address:
  Dr. Paul D. Siegel, Allergy and Clinical Immunology Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, 1095 Willowdale Road, Morgantown, WV 26505-2888
• Email:
  pds3@cdc.gov
• CAS Registry Number:
  Toluene 2,4-diisocyanate (CAS RN 584-84-9)
• Federal Fiscal Year:
  2010
• NORA Priority Area:
  Healthcare and Social Assistance ; Services
• Peer Reviewed:
  True
• Source Full Name:
  Hybridoma
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:67dfc89233f5affefc8acf2d77b29ea4000ccd3fb37af12b8d6d665418abc35da8dc341f9b9a96a7916bfc339b30cf470f6ceeea91c2a309ddc0a6dc8fe1f970
• Download URL:
  https://stacks.cdc.gov/view/cdc/188215/cdc_188215_DS1.pdf
• File Type:
  [PDF - 813.18 KB ]