Allergen binding to specific IgE in the nasal mucosa of allergic patients  Alex KleinJan, BSca, Tom Godthelp, MDa, Albert van Toornenenbergen, PhDb, Wytske J. Fokkens, MD, PhDa  Journal of Allergy and Clinical Immunology  Volume 99, Issue 4, Pages 515-521 (April 1997) DOI: 10.1016/S0091-6749(97)70079-4 Copyright © 1997 Mosby, Inc. Terms and Conditions

Figure 1 Number of allergen-positive cells in epithelium (A) and lamina propria (B). Staining with biotinylated grass pollen (GP1) showed positive cells in patients with grass pollen allergy only. Staining with biotinylated D. pteronyssinus (D1) only showed positive cells in patients with perennial allergy. Journal of Allergy and Clinical Immunology 1997 99, 515-521DOI: (10.1016/S0091-6749(97)70079-4) Copyright © 1997 Mosby, Inc. Terms and Conditions

Figure 1 Number of allergen-positive cells in epithelium (A) and lamina propria (B). Staining with biotinylated grass pollen (GP1) showed positive cells in patients with grass pollen allergy only. Staining with biotinylated D. pteronyssinus (D1) only showed positive cells in patients with perennial allergy. Journal of Allergy and Clinical Immunology 1997 99, 515-521DOI: (10.1016/S0091-6749(97)70079-4) Copyright © 1997 Mosby, Inc. Terms and Conditions

Figure 2 Blocking experiment by preincubation with nonbiotinylated allergens. Blocking experiments proved the method to be highly specific. Journal of Allergy and Clinical Immunology 1997 99, 515-521DOI: (10.1016/S0091-6749(97)70079-4) Copyright © 1997 Mosby, Inc. Terms and Conditions

Figure 3 Cryostat section of nasal mucosa of a patient with isolated grass pollen allergy (A, C, F) and a patient with perennial allergy to HDM (B, D, E). A, Single staining with GP1 in patient allergic to grass pollen; allergen-positive cells stained red. B, Single staining with GP1 in patient allergic to HDM; no GP1-positive cells were observed. Double-staining allergen (GP1 or D1) and mouse monoclonal antibodies against: IgE (C), HLA-DR (D), mast cell chymase (E), and Langerhans cells (CD1) (F). Allergen-positive cells were sharply localized, red-staining cells (arrowheads). Cells staining positive for a monoclonal antibody stained soft blue (open arrows); double-positive cells were a mixed color and dark (filled arrows). Allergen-positive cells stained double with IgE, FcϵRI, CD1, HLA-DR, tryptase, and chymase. Most allergen-positive cells proved to be mast cells. Journal of Allergy and Clinical Immunology 1997 99, 515-521DOI: (10.1016/S0091-6749(97)70079-4) Copyright © 1997 Mosby, Inc. Terms and Conditions

Figure 3 Cryostat section of nasal mucosa of a patient with isolated grass pollen allergy (A, C, F) and a patient with perennial allergy to HDM (B, D, E). A, Single staining with GP1 in patient allergic to grass pollen; allergen-positive cells stained red. B, Single staining with GP1 in patient allergic to HDM; no GP1-positive cells were observed. Double-staining allergen (GP1 or D1) and mouse monoclonal antibodies against: IgE (C), HLA-DR (D), mast cell chymase (E), and Langerhans cells (CD1) (F). Allergen-positive cells were sharply localized, red-staining cells (arrowheads). Cells staining positive for a monoclonal antibody stained soft blue (open arrows); double-positive cells were a mixed color and dark (filled arrows). Allergen-positive cells stained double with IgE, FcϵRI, CD1, HLA-DR, tryptase, and chymase. Most allergen-positive cells proved to be mast cells. Journal of Allergy and Clinical Immunology 1997 99, 515-521DOI: (10.1016/S0091-6749(97)70079-4) Copyright © 1997 Mosby, Inc. Terms and Conditions

Figure 3 Cryostat section of nasal mucosa of a patient with isolated grass pollen allergy (A, C, F) and a patient with perennial allergy to HDM (B, D, E). A, Single staining with GP1 in patient allergic to grass pollen; allergen-positive cells stained red. B, Single staining with GP1 in patient allergic to HDM; no GP1-positive cells were observed. Double-staining allergen (GP1 or D1) and mouse monoclonal antibodies against: IgE (C), HLA-DR (D), mast cell chymase (E), and Langerhans cells (CD1) (F). Allergen-positive cells were sharply localized, red-staining cells (arrowheads). Cells staining positive for a monoclonal antibody stained soft blue (open arrows); double-positive cells were a mixed color and dark (filled arrows). Allergen-positive cells stained double with IgE, FcϵRI, CD1, HLA-DR, tryptase, and chymase. Most allergen-positive cells proved to be mast cells. Journal of Allergy and Clinical Immunology 1997 99, 515-521DOI: (10.1016/S0091-6749(97)70079-4) Copyright © 1997 Mosby, Inc. Terms and Conditions

Figure 3 Cryostat section of nasal mucosa of a patient with isolated grass pollen allergy (A, C, F) and a patient with perennial allergy to HDM (B, D, E). A, Single staining with GP1 in patient allergic to grass pollen; allergen-positive cells stained red. B, Single staining with GP1 in patient allergic to HDM; no GP1-positive cells were observed. Double-staining allergen (GP1 or D1) and mouse monoclonal antibodies against: IgE (C), HLA-DR (D), mast cell chymase (E), and Langerhans cells (CD1) (F). Allergen-positive cells were sharply localized, red-staining cells (arrowheads). Cells staining positive for a monoclonal antibody stained soft blue (open arrows); double-positive cells were a mixed color and dark (filled arrows). Allergen-positive cells stained double with IgE, FcϵRI, CD1, HLA-DR, tryptase, and chymase. Most allergen-positive cells proved to be mast cells. Journal of Allergy and Clinical Immunology 1997 99, 515-521DOI: (10.1016/S0091-6749(97)70079-4) Copyright © 1997 Mosby, Inc. Terms and Conditions

Figure 3 Cryostat section of nasal mucosa of a patient with isolated grass pollen allergy (A, C, F) and a patient with perennial allergy to HDM (B, D, E). A, Single staining with GP1 in patient allergic to grass pollen; allergen-positive cells stained red. B, Single staining with GP1 in patient allergic to HDM; no GP1-positive cells were observed. Double-staining allergen (GP1 or D1) and mouse monoclonal antibodies against: IgE (C), HLA-DR (D), mast cell chymase (E), and Langerhans cells (CD1) (F). Allergen-positive cells were sharply localized, red-staining cells (arrowheads). Cells staining positive for a monoclonal antibody stained soft blue (open arrows); double-positive cells were a mixed color and dark (filled arrows). Allergen-positive cells stained double with IgE, FcϵRI, CD1, HLA-DR, tryptase, and chymase. Most allergen-positive cells proved to be mast cells. Journal of Allergy and Clinical Immunology 1997 99, 515-521DOI: (10.1016/S0091-6749(97)70079-4) Copyright © 1997 Mosby, Inc. Terms and Conditions

Figure 3 Cryostat section of nasal mucosa of a patient with isolated grass pollen allergy (A, C, F) and a patient with perennial allergy to HDM (B, D, E). A, Single staining with GP1 in patient allergic to grass pollen; allergen-positive cells stained red. B, Single staining with GP1 in patient allergic to HDM; no GP1-positive cells were observed. Double-staining allergen (GP1 or D1) and mouse monoclonal antibodies against: IgE (C), HLA-DR (D), mast cell chymase (E), and Langerhans cells (CD1) (F). Allergen-positive cells were sharply localized, red-staining cells (arrowheads). Cells staining positive for a monoclonal antibody stained soft blue (open arrows); double-positive cells were a mixed color and dark (filled arrows). Allergen-positive cells stained double with IgE, FcϵRI, CD1, HLA-DR, tryptase, and chymase. Most allergen-positive cells proved to be mast cells. Journal of Allergy and Clinical Immunology 1997 99, 515-521DOI: (10.1016/S0091-6749(97)70079-4) Copyright © 1997 Mosby, Inc. Terms and Conditions