Immunofluorescent staining of MBP in MCs and eosinophils.

Confocal fluorescence image with anti-MBP MoAb (A) and differential interference contrast image (B) of permeabilized CB-derived cultured MCs (1-μm section). Confocal fluorescence images with anti-MBP MoAb of permeabilized lung-derived purified MCs (C) and permeabilized PB-derived purified eosinophils (D). MBP was also detected even in unpermeabilized CB-derived cultured MCs, but only after anti-IgE challenge. (E) Confocal fluorescence and differential interference contrast images were synthesized in the figure. (F) Double staining for MBP (FITC) and tryptase (rhodamine) of a skin biopsy sample obtained from a patient with severe atopic dermatitis. Green images represent MBP-only positive cells, whereas orange-yellow images show MBP and tryptase double-positive cells. Tryptase-only positive cells (red images) were not found in this study, including samples from 2 patients with atopic dermatitis and 2 normal control subjects. Use of a control antibody confirmed all of the positive staining findings to be specific for anti-MBP and antitryptase antibodies.