Molecular determinants of humoral immune specificity for the occupational allergen, methylene diphenyl diisocyanate

Mol Immunol. 2013 Jun;54(2):233-7. doi: 10.1016/j.molimm.2012.11.017. Epub 2013 Jan 4.


Methylene diphenyl diisocyanate (MDI), a low molecular weight chemical important for producing polyurethane foam, coatings, and elastomers is a major cause of occupational asthma, however, mechanisms of disease pathogenesis remain poorly understood. This study characterizes the rearranged germline and hypervariable region cDNA of new anti-MDI secreting hybridomas derived from mice immunized with MDI-conjugated to autologous serum proteins. Six IgG1 secreting clones were identified in initial screening ELISAs, based on differential binding to MDI conjugated human albumin vs. mock exposed albumin. The mAbs secreted by the hybridomas also recognized MDI conjugated to other model proteins (e.g. ovalbumin, transferrin), but did not bind unconjugated proteins, or protein conjugates prepared with other isocyanates (e.g. TDI, HDI). The mAbs displayed MDI-dose dependent binding in ELISA and Western blot, and exhibited varying degrees of cross-competition, suggesting differences in epitope specificity. The cDNA encoding the monoclonal antibodies reveal clonal differences in the CDR3 regions, germline gene usage, and patterns of somatic hypermutation related to epitope specificity. Together, the data provide new insight into the molecular determinants of humoral MDI specificity, and characterize anti-MDI IgG1 mAbs that may be developed into useful diagnostic reagents.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Allergens / immunology*
  • Allergens / metabolism
  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / metabolism
  • Antibody Specificity / immunology
  • Binding, Competitive / immunology
  • Cross Reactions / immunology
  • Female
  • Humans
  • Hybridomas / immunology
  • Hybridomas / metabolism
  • Immunity, Humoral*
  • Immunoglobulin G / chemistry
  • Immunoglobulin G / immunology
  • Immunoglobulin G / metabolism
  • Isocyanates / immunology*
  • Isocyanates / metabolism
  • Mice
  • Occupational Exposure
  • Protein Binding / immunology


  • Allergens
  • Antibodies, Monoclonal
  • Immunoglobulin G
  • Isocyanates
  • 4,4'-diphenylmethane diisocyanate