Allergy to rabbits. I. Specificity and non-specificity of RAST and crossed-radioimmunoelectrophoresis due to the presence of light chains in rabbit allergenic extracts.
Investigations have been carried out into the presence of antibody light chains in rabbit allergenic extracts and the interference in RAST and crossed-radioimmunoelectrophoresis (XRIE) caused by antibodies directed against them. A "non-specific" uptake of radioactivity in XRIE has been demonstrated to be caused by direct cross-linking of the 125I rabbit anti-human IgE by the sheep antibodies in the immunoprecipitate of rabbit light chains. Preincubation with normal rabbit serum blocked this direct uptake of the labelled antibody and enabled specific IgE uptake on the light chains to be demonstrated for rabbit allergic sera. Verification of the allergenicity of the light chains was obtained from a specific light chain RAST. Elution from a Sephacryl S-200 gel filtration column indicated a MW of approx. 50 Kd and confirmation of the component as light chain dimers, not Fab fragments, was obtained by allotyping for loci present on heavy chains and light chains in the Fab region. Light chains were detected in urine from rabbits of all ages and in an extract of dust collected in a rabbit housing area. No background staining was observed in XRIE using rabbit antisera, either with rabbit allergic sera with specific IgE or with a human serum containing specific IgG antibodies to rabbit IgG. This latter serum also showed no evidence of uptake on all immunoprecipitates in systems using rabbit antisera, and did not give false positive RAST results when the labelled rabbit anti-human IgE contained unlabelled rabbit IgG. Those sera with specific IgE to light chains showed no uptake in XRIE using rabbit antisera, indicating that the IgE was possibly specific for epitopes revealed by the dissociation of the whole IgG molecule.
D000485:Allergens / Q000302:isolation & purification*; D000818:Animals; D000918:Antibody Specificity; D006801:Humans; D006967:Hypersensitivity / Q000209:etiology*; D007123:Immunoelectrophoresis, Two-Dimensional; D007073:Immunoglobulin E / Q000032:analysis; D007074:Immunoglobulin G / Q000302:isolation & purification; D007147:Immunoglobulin Light Chains / Q000302:isolation & purification*; D011487:Protein Conformation; D011817:Rabbits / Q000276:immunology*; D011852:Radioallergosorbent Test
D000485:Allergens; D007074:Immunoglobulin G; D007147:Immunoglobulin Light Chains; D007073:Immunoglobulin E