Isolation of dinitrophenyl-skin protein conjugates by immunoadsorbent chromatography.
Immunoadsorbent columns charged with rabbit anti-dinitrophenyl (DNP) antibody were used to isolate and concentrate DNP-protein conjugates from the 105,000 g supernate of extracts prepared from guinea pig epidermis previously painted with dinitrofluorobenzene (DNFB). Conjugates isolated in this manner elicited positive delayed hypersensitivity skin tests when injected intradermally into DNFB-sensitized animals. Dinitrophenylated epidermal protein conjugates prepared in vitro were capable of specifically inhibiting the migration of peritoneal exudate cells obtained from DNFB-sensitized guinea pigs. These results demonstrate the application of immunoadsorbent chromatography and the migration inhibition assay to the further definition of the epitopes involved in contact sensitivity.
D000818:Animals; D002464:Cell Migration Inhibition; D003877:Dermatitis, Contact / Q000276:immunology; D004136:Dinitrobenzenes / Q000276:immunology* / Q000302:isolation & purification; D005260:Female; D006168:Guinea Pigs; D006968:Hypersensitivity, Delayed / Q000276:immunology*; D007163:Immunosorbent Techniques; D008264:Macrophages / Q000276:immunology; D009578:Nitrobenzenes / Q000276:immunology*; D011506:Proteins / Q000276:immunology* / Q000302:isolation & purification; D012867:Skin / Q000276:immunology*; D012882:Skin Tests
D004136:Dinitrobenzenes; D009578:Nitrobenzenes; D011506:Proteins