Immunochemical studies on red cell auto antigens: use and limits of immunoprecipitation from biotinylated erythrocyte membrane.
Erythrocyte surface was labelled by means of biotin; immunoprecipitation technique was then used to localise antigens recognised on red cell membrane proteins by: a) autoantibodies from 13 patients with antierythrocyte autoimmunity; b) commercially available anti-D and anti-k (Cellano) antierythrocyte alloantibodies. Results with alloantibodies are comparable to those obtained using radiochemical probes. Immunoprecipitations with autoantibody containing eluates showed reactivity at different molecular weights (the most common at 34-50 kD, others at 100 and 45 kD and a newly described one at 80 kD), thus confirming that many membrane proteins may act as target antigens for erythrocyte autoimmunity. We found a higher percentage of reactive immunoprecipitates than previously reported using the same labelling method. However, critical conditions to allow valuable results seem to be a threshold amount of autoantibody to precipitate any recognisable band and the sensitivity of the detection method. Hence methodological variables must be taken into consideration before concluding that "non protein" antigens trigger the autoimmune process.
De Angelis
V
V
Servizio Immunotrasfusionale e Analisi Cliniche, IRCCS Centro di Riferimento Oncologico, Aviano, Italy.
Pradella
P
P
Biasinutto
C
C
Steffan
A
A
Cozzi
M R
MR
D001324:Autoantigens / Q000032:analysis*; D001710:Biotin; D004910:Erythrocyte Membrane / Q000276:immunology*; D006801:Humans; D011233:Precipitin Tests
D001324:Autoantigens; D001710:Biotin