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Predominance of type 2 cytokine–producing CD4+ and CD8+ cells in patients with atopic dermatitis  Masatoshi Nakazawa, DVMa, Nakako Sugi, MDb, Hiroshi.

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Presentation on theme: "Predominance of type 2 cytokine–producing CD4+ and CD8+ cells in patients with atopic dermatitis  Masatoshi Nakazawa, DVMa, Nakako Sugi, MDb, Hiroshi."— Presentation transcript:

1 Predominance of type 2 cytokine–producing CD4+ and CD8+ cells in patients with atopic dermatitis 
Masatoshi Nakazawa, DVMa, Nakako Sugi, MDb, Hiroshi Kawaguchi, MD, PhDc, Norihisa Ishii, MD, PhDc, Hiroshi Nakajima, MD, PhDc, Mutsuhiko Minami, MD, PhDa  Journal of Allergy and Clinical Immunology  Volume 99, Issue 5, Pages (May 1997) DOI: /S (97) Copyright © 1997 Mosby, Inc. Terms and Conditions

2 Fig. 1 PBMCs from a patient with AD were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-2 (A) or IL-4 (B), and CD4. An analysis gate was set on lymphocytes, by forward- and side-scatter properties, and CD4+ cells. The histograms illustrate intracellular staining obtained in the presence (solid line) or absence (shaded) of excess amounts of specific recombinant cytokines. Intensity of fluorescence blocked by excess amounts of recombinant cytokines was overlapped with intensity of fluorescence of the negative control (dotted line). Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

3 Fig. 1 PBMCs from a patient with AD were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-2 (A) or IL-4 (B), and CD4. An analysis gate was set on lymphocytes, by forward- and side-scatter properties, and CD4+ cells. The histograms illustrate intracellular staining obtained in the presence (solid line) or absence (shaded) of excess amounts of specific recombinant cytokines. Intensity of fluorescence blocked by excess amounts of recombinant cytokines was overlapped with intensity of fluorescence of the negative control (dotted line). Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

4 Fig. 2 PBMCs from patients with AD were stimulated for 6 or 24 hours with immobilized anti-CD3 mAb, and monensin was added for the last 6 hours. Cells were stained for IL-4, IL-2, or IFN-γ. Frequencies of cells producing each cytokine were examined at 0 hours (without stimulation) and after 6 and 24 hours of stimulation with anti-CD3 mAb. Frequency of cytokine-producing cells is shown as the mean percentage ± SD of the CD4+ (A) or the CD8+ cell population (B). Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

5 Fig. 2 PBMCs from patients with AD were stimulated for 6 or 24 hours with immobilized anti-CD3 mAb, and monensin was added for the last 6 hours. Cells were stained for IL-4, IL-2, or IFN-γ. Frequencies of cells producing each cytokine were examined at 0 hours (without stimulation) and after 6 and 24 hours of stimulation with anti-CD3 mAb. Frequency of cytokine-producing cells is shown as the mean percentage ± SD of the CD4+ (A) or the CD8+ cell population (B). Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

6 Fig. 3 PBMCs from patients with AD, disease controls (patients with chronic psoriasis [circles with diagonal lines] or contact dermatitis [shaded circles]), and healthy control subjects were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-4 (A and B), IL-2 (C and D), or IFN-γ (E and F). Frequencies of cells producing each cytokine were examined at 0 hours (without stimulation) (A, C, and E) and after 6 hours of stimulation with anti-CD3 mAb (B, D, and F). Frequency of cytokine-producing cells is shown as a percentage of the CD4+ cell population. Lines (|pk) show mean ± SD. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

7 Fig. 3 PBMCs from patients with AD, disease controls (patients with chronic psoriasis [circles with diagonal lines] or contact dermatitis [shaded circles]), and healthy control subjects were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-4 (A and B), IL-2 (C and D), or IFN-γ (E and F). Frequencies of cells producing each cytokine were examined at 0 hours (without stimulation) (A, C, and E) and after 6 hours of stimulation with anti-CD3 mAb (B, D, and F). Frequency of cytokine-producing cells is shown as a percentage of the CD4+ cell population. Lines (|pk) show mean ± SD. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

8 Fig. 3 PBMCs from patients with AD, disease controls (patients with chronic psoriasis [circles with diagonal lines] or contact dermatitis [shaded circles]), and healthy control subjects were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-4 (A and B), IL-2 (C and D), or IFN-γ (E and F). Frequencies of cells producing each cytokine were examined at 0 hours (without stimulation) (A, C, and E) and after 6 hours of stimulation with anti-CD3 mAb (B, D, and F). Frequency of cytokine-producing cells is shown as a percentage of the CD4+ cell population. Lines (|pk) show mean ± SD. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

9 Fig. 3 PBMCs from patients with AD, disease controls (patients with chronic psoriasis [circles with diagonal lines] or contact dermatitis [shaded circles]), and healthy control subjects were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-4 (A and B), IL-2 (C and D), or IFN-γ (E and F). Frequencies of cells producing each cytokine were examined at 0 hours (without stimulation) (A, C, and E) and after 6 hours of stimulation with anti-CD3 mAb (B, D, and F). Frequency of cytokine-producing cells is shown as a percentage of the CD4+ cell population. Lines (|pk) show mean ± SD. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

10 Fig. 3 PBMCs from patients with AD, disease controls (patients with chronic psoriasis [circles with diagonal lines] or contact dermatitis [shaded circles]), and healthy control subjects were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-4 (A and B), IL-2 (C and D), or IFN-γ (E and F). Frequencies of cells producing each cytokine were examined at 0 hours (without stimulation) (A, C, and E) and after 6 hours of stimulation with anti-CD3 mAb (B, D, and F). Frequency of cytokine-producing cells is shown as a percentage of the CD4+ cell population. Lines (|pk) show mean ± SD. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

11 Fig. 3 PBMCs from patients with AD, disease controls (patients with chronic psoriasis [circles with diagonal lines] or contact dermatitis [shaded circles]), and healthy control subjects were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-4 (A and B), IL-2 (C and D), or IFN-γ (E and F). Frequencies of cells producing each cytokine were examined at 0 hours (without stimulation) (A, C, and E) and after 6 hours of stimulation with anti-CD3 mAb (B, D, and F). Frequency of cytokine-producing cells is shown as a percentage of the CD4+ cell population. Lines (|pk) show mean ± SD. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

12 Fig. 4 PBMCs from patients with AD, disease controls (patients with chronic psoriasis [circles with diagonal lines] or contact dermatitis [shaded circles]), and healthy control subjects were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-4 (A and B), IL-2 (C and D), or IFN-γ (E and F). Frequencies of cells producing each cytokine were examined at 0 hours (without stimulation) (A, C, and E) and after 6 hours of stimulation with anti-CD3 mAb (B, D, and F). Frequency of cytokine-producing cells is shown as a percentage of the CD8+ cell population. Lines (|pk) show mean ± SD. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

13 Fig. 4 PBMCs from patients with AD, disease controls (patients with chronic psoriasis [circles with diagonal lines] or contact dermatitis [shaded circles]), and healthy control subjects were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-4 (A and B), IL-2 (C and D), or IFN-γ (E and F). Frequencies of cells producing each cytokine were examined at 0 hours (without stimulation) (A, C, and E) and after 6 hours of stimulation with anti-CD3 mAb (B, D, and F). Frequency of cytokine-producing cells is shown as a percentage of the CD8+ cell population. Lines (|pk) show mean ± SD. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

14 Fig. 4 PBMCs from patients with AD, disease controls (patients with chronic psoriasis [circles with diagonal lines] or contact dermatitis [shaded circles]), and healthy control subjects were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-4 (A and B), IL-2 (C and D), or IFN-γ (E and F). Frequencies of cells producing each cytokine were examined at 0 hours (without stimulation) (A, C, and E) and after 6 hours of stimulation with anti-CD3 mAb (B, D, and F). Frequency of cytokine-producing cells is shown as a percentage of the CD8+ cell population. Lines (|pk) show mean ± SD. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

15 Fig. 4 PBMCs from patients with AD, disease controls (patients with chronic psoriasis [circles with diagonal lines] or contact dermatitis [shaded circles]), and healthy control subjects were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-4 (A and B), IL-2 (C and D), or IFN-γ (E and F). Frequencies of cells producing each cytokine were examined at 0 hours (without stimulation) (A, C, and E) and after 6 hours of stimulation with anti-CD3 mAb (B, D, and F). Frequency of cytokine-producing cells is shown as a percentage of the CD8+ cell population. Lines (|pk) show mean ± SD. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

16 Fig. 4 PBMCs from patients with AD, disease controls (patients with chronic psoriasis [circles with diagonal lines] or contact dermatitis [shaded circles]), and healthy control subjects were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-4 (A and B), IL-2 (C and D), or IFN-γ (E and F). Frequencies of cells producing each cytokine were examined at 0 hours (without stimulation) (A, C, and E) and after 6 hours of stimulation with anti-CD3 mAb (B, D, and F). Frequency of cytokine-producing cells is shown as a percentage of the CD8+ cell population. Lines (|pk) show mean ± SD. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

17 Fig. 4 PBMCs from patients with AD, disease controls (patients with chronic psoriasis [circles with diagonal lines] or contact dermatitis [shaded circles]), and healthy control subjects were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-4 (A and B), IL-2 (C and D), or IFN-γ (E and F). Frequencies of cells producing each cytokine were examined at 0 hours (without stimulation) (A, C, and E) and after 6 hours of stimulation with anti-CD3 mAb (B, D, and F). Frequency of cytokine-producing cells is shown as a percentage of the CD8+ cell population. Lines (|pk) show mean ± SD. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

18 Fig. 5 PBMCs from patients with AD or healthy control subjects were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-4 (A and B), IL-2 (C and D), or IFN-γ (E and F). Frequency of cytokine-producing cells is shown as a percentage of the CD4+ cell (A, C, and E) or CD8+ cell (B, D, and F) population. Patients with AD were divided into three groups according to disease severity. Lines (|pk) show mean ± SD. *Frequency of IL-4–producing CD4+ and CD8+ cells from healthy control subjects was significantly lower (p < 0.001) than that from any group of patients with AD. **Frequency of IFN-γ–producing CD4+ and CD8+ cells from healthy control subjects was significantly higher (p < 0.01) than that from any group of patients with AD. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

19 Fig. 5 PBMCs from patients with AD or healthy control subjects were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-4 (A and B), IL-2 (C and D), or IFN-γ (E and F). Frequency of cytokine-producing cells is shown as a percentage of the CD4+ cell (A, C, and E) or CD8+ cell (B, D, and F) population. Patients with AD were divided into three groups according to disease severity. Lines (|pk) show mean ± SD. *Frequency of IL-4–producing CD4+ and CD8+ cells from healthy control subjects was significantly lower (p < 0.001) than that from any group of patients with AD. **Frequency of IFN-γ–producing CD4+ and CD8+ cells from healthy control subjects was significantly higher (p < 0.01) than that from any group of patients with AD. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

20 Fig. 5 PBMCs from patients with AD or healthy control subjects were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-4 (A and B), IL-2 (C and D), or IFN-γ (E and F). Frequency of cytokine-producing cells is shown as a percentage of the CD4+ cell (A, C, and E) or CD8+ cell (B, D, and F) population. Patients with AD were divided into three groups according to disease severity. Lines (|pk) show mean ± SD. *Frequency of IL-4–producing CD4+ and CD8+ cells from healthy control subjects was significantly lower (p < 0.001) than that from any group of patients with AD. **Frequency of IFN-γ–producing CD4+ and CD8+ cells from healthy control subjects was significantly higher (p < 0.01) than that from any group of patients with AD. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

21 Fig. 5 PBMCs from patients with AD or healthy control subjects were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-4 (A and B), IL-2 (C and D), or IFN-γ (E and F). Frequency of cytokine-producing cells is shown as a percentage of the CD4+ cell (A, C, and E) or CD8+ cell (B, D, and F) population. Patients with AD were divided into three groups according to disease severity. Lines (|pk) show mean ± SD. *Frequency of IL-4–producing CD4+ and CD8+ cells from healthy control subjects was significantly lower (p < 0.001) than that from any group of patients with AD. **Frequency of IFN-γ–producing CD4+ and CD8+ cells from healthy control subjects was significantly higher (p < 0.01) than that from any group of patients with AD. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

22 Fig. 5 PBMCs from patients with AD or healthy control subjects were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-4 (A and B), IL-2 (C and D), or IFN-γ (E and F). Frequency of cytokine-producing cells is shown as a percentage of the CD4+ cell (A, C, and E) or CD8+ cell (B, D, and F) population. Patients with AD were divided into three groups according to disease severity. Lines (|pk) show mean ± SD. *Frequency of IL-4–producing CD4+ and CD8+ cells from healthy control subjects was significantly lower (p < 0.001) than that from any group of patients with AD. **Frequency of IFN-γ–producing CD4+ and CD8+ cells from healthy control subjects was significantly higher (p < 0.01) than that from any group of patients with AD. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

23 Fig. 5 PBMCs from patients with AD or healthy control subjects were stimulated for 6 hours with immobilized anti-CD3 mAb in the presence of monensin. Cells were stained for IL-4 (A and B), IL-2 (C and D), or IFN-γ (E and F). Frequency of cytokine-producing cells is shown as a percentage of the CD4+ cell (A, C, and E) or CD8+ cell (B, D, and F) population. Patients with AD were divided into three groups according to disease severity. Lines (|pk) show mean ± SD. *Frequency of IL-4–producing CD4+ and CD8+ cells from healthy control subjects was significantly lower (p < 0.001) than that from any group of patients with AD. **Frequency of IFN-γ–producing CD4+ and CD8+ cells from healthy control subjects was significantly higher (p < 0.01) than that from any group of patients with AD. Journal of Allergy and Clinical Immunology  , DOI: ( /S (97) ) Copyright © 1997 Mosby, Inc. Terms and Conditions

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