Harald Renz, MD, Chaya Brodie, PhD, Katherine Bradley, BS, Donald Y. M

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Presentation transcript:

Enhancement of IgE production by anti-CD40 antibody in atopic dermatitis Harald Renz, MD, Chaya Brodie, PhD, Katherine Bradley, BS, Donald Y.M. Leung, MD, PhD, Erwin W. Gelfand, MD Journal of Allergy and Clinical Immunology Volume 93, Issue 3, Pages 658-668 (March 1994) DOI: 10.1016/S0091-6749(94)70078-8 Copyright © 1994 Mosby, Inc. Terms and Conditions

FIG. 1 Induction of IgE production after incubation with anti-CD40 or the combination of anti-CD40 and IL-4. PBMCs from five atopic and five nonatopic donors were cultured with anti-CD40 (0.5 μg/ml), IL-4 (400 U/ml), or tissue culture medium alone for 14 days. IgE production was then analyzed in culture supernatants by ELISA. In each individual the effect of anti-CD40 stimulation was compared with control levels (culture with medium alone) and calculated as percent control (means: atopic donors, 4536 pg/ml; normal donors, 508 pg/ml). Similarly, in IL-4–stimulated cultures the effect of anti-CD40 was compared with cultures treated with IL-4 alone (control) and expressed as percent control (means: atopic donors, 11,144 pg/ml; normal donors, 4046 pg/ml). For each individual, cultures were performed in duplicate. Shown are means ± SD for each experimental group. The asterisk indicates significant (p < 0.05) differences between the groups. Journal of Allergy and Clinical Immunology 1994 93, 658-668DOI: (10.1016/S0091-6749(94)70078-8) Copyright © 1994 Mosby, Inc. Terms and Conditions

FIG. 2 Effect of anti-CD40 treatment on IgG, IgM, and IgA production. PBMCs from five atopic and five nonatopic donors were cultured with anti-CD40 (0.5 μg/ml) ± IL-4 (400 U/ml) for 14 days, and Ig isotype production was measured in culture supernatants by ELISA. The effect of anti-CD40 was expressed as percent control (cultures with tissue culture medium alone). The baseline levels were: for atopic donors, IgM 868 ng/ml, IgG 926 ng/ml, IgA 95 ng/ml; for normal donors, IgM 716 ng/ml, IgG 672 ng/ml, IgA 104 ng/ml. Shown are means ± SD for each experimental group. NS, Not significant. Journal of Allergy and Clinical Immunology 1994 93, 658-668DOI: (10.1016/S0091-6749(94)70078-8) Copyright © 1994 Mosby, Inc. Terms and Conditions

FIG. 3 Lymphocyte proliferation induced by anti-IgM, anti-CD40, and IL-4. PBMCs from five atopic patients and five control donors were incubated with anti-IgM (0.5 μg/ml), anti-CD40 (500 ng/ml), and IL-4 (400 U/ml) in different combinations as shown in the figure. The cell cultures were carried out for 72 hours. During the last 6 hours of the culture period tritiated thymidine (TdR) was added (1 μCi/well). In each donor the proliferative response to the reagents was compared with the medium control, and the effect was expressed as percent of the medium control (100%). Expressed are means ± SD from all donors in each group. Mean counts per minute of the medium control were 1345 cpm in the control group and 1947 cpm in the group of atopic patients. Journal of Allergy and Clinical Immunology 1994 93, 658-668DOI: (10.1016/S0091-6749(94)70078-8) Copyright © 1994 Mosby, Inc. Terms and Conditions

FIG. 4 Analysis of CD40 and CD3 expression on lymphocytes from one normal nonatopic donor (N) and one patient with atopic dermatitis (AD) by double staining. Indicated are MFI and the percentage of CD40-positive cells. Cells were analyzed with a gate for lymphocytes. Journal of Allergy and Clinical Immunology 1994 93, 658-668DOI: (10.1016/S0091-6749(94)70078-8) Copyright © 1994 Mosby, Inc. Terms and Conditions

FIG. 5 Regulation of CD40 expression by IL-4. PBMCs from two normal donors were incubated with either IL-4 or medium alone for 14 days. At the end of the culture period, lymphocytes were recovered and stained with anti-CD40 as described in the Methods section. Shown are the percentage of CD-40-positive cells and the MFI. The percent of CD-40–positive cells was identical to the percent of CD20-expressing cells (data not shown). Journal of Allergy and Clinical Immunology 1994 93, 658-668DOI: (10.1016/S0091-6749(94)70078-8) Copyright © 1994 Mosby, Inc. Terms and Conditions