Immunofluorescence studies on eosinophilic granulocytes.
Immunofluorescence techniques were used to find out whether immunoglobulins (Ig) and complement (C3) determinants are present on the membrane of eosinophilic granulocytes in atopic patients and in patients with an allergic contact dermatitis. Normal healthy individuals served as controls. The studies were performed with eosinophilic granulocytes in suspension and in thin cell layers. The demonstration of the presence of cytoplasmic Ig and C3 in eosinophilic granulocytes was carried out in skin specimens taken 20-30 min after injection of the antigen and in specimens taken from positive patch tests 48 h after application of the antigen. No immunoglobulins or C3 determinants could be demonstrated on the membrane of eosinophilic granulocytes in the patients or the healthy controls, when the suspension methods were used. However, in thin cell layers and in skin sections the eosinophilic granulocytes showed a positive fluorescence of the cytoplasm after having been exposed to various conjugates and a FITC solution. It is concluded that the commonly used immunofluorescence techniques tend to give non-specific staining results, most likely due to reactivity of the FITC marker of the conjugates with basic proteins present in the cytoplasm of the eosinophilic granulocyte.
D003176:Complement C3 / Q000032:analysis; D003593:Cytoplasm / Q000276:immunology; D003876:Dermatitis, Atopic / Q000276:immunology*; D003877:Dermatitis, Contact / Q000276:immunology*; D004804:Eosinophils / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D006801:Humans; D011947:Receptors, Antigen, B-Cell / Q000032:analysis; D012867:Skin / Q000276:immunology
D003176:Complement C3; D011947:Receptors, Antigen, B-Cell