Delayed eosinophil programmed cell death in vitro: A common feature of inhalant allergy and extrinsic and intrinsic atopic dermatitis Bettina Wedi, MD,

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Presentation transcript:

Delayed eosinophil programmed cell death in vitro: A common feature of inhalant allergy and extrinsic and intrinsic atopic dermatitis Bettina Wedi, MD, Ulrike Raap, Heitje Lewrick, Alexander Kapp, MD Journal of Allergy and Clinical Immunology Volume 100, Issue 4, Pages 536-543 (October 1997) DOI: 10.1016/S0091-6749(97)70147-7 Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 1 Comparison of survival over time of eosinophils from subjects with AD, subjects with IA, and NA subjects. Culture was in medium or with cytokines (100 U/ml each). The data are given as mean ± SEM values of 4 to 11 separate experiments with eosinophils of different patients. Survival of unstimulated eosinophils (med) was significantly increased at all time-points studied in IA compared to NA subjects (p < 0.05), in AD compared to NA subjects (p < 0.0001), and in AD compared to IA (p < 0.05) (not indicated). *, p < 0.05; **, p < 0.01; ***, p < 0.0001 (stimulus versus medium). Journal of Allergy and Clinical Immunology 1997 100, 536-543DOI: (10.1016/S0091-6749(97)70147-7) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 2 DNA was extracted from eosinophils of a NA subject (lanes 2 to 6), a patient with IA (lanes 7 to 11), and a patient with AD (lanes 12 to 16) as described in Material and Methods. Lane 1 contains DNA standard made by HindIII-digested lambda DNA. After electrophoresis of the DNA in 1.8% agarose and staining with ethidium bromide, the DNA was viewed by UV fluorescence. After 48 to 72 hours, DNA extracted from the NA subject but not DNA from the patient with IA or the patient with AD exhibited a typical ladder consisting of characteristic 200 base pair DNA fragments. In IA, PCD occurred after 96 hours, whereas in AD, PCD was not evident up to 144 hours. One representative example is shown. Hours in culture are indicated above the photograph. Journal of Allergy and Clinical Immunology 1997 100, 536-543DOI: (10.1016/S0091-6749(97)70147-7) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 3 Detection of proeosinophilic cytokines released into the eosinophil supernatants of subjects with inhalant allergy (IA) or atopic dermatitis (AD) and nonatopic subjects (NA), as assessed by specific enzyme immunoassays. Data are given as mean ± SEM values of 4 to 10 different donors in each group. Unfilled bars represent supernatants obtained after 72 hours of culture; filled bars represent supernatants obtained after 144 hours in culture. The minimum detectable concentration for GM-CSF and IL-5 was 7.8 pg/ml. A, Detection of GM-CSF. B, Detection of IL-5. *p = 0.006 versus NA and p < 0.05 versus IA. Journal of Allergy and Clinical Immunology 1997 100, 536-543DOI: (10.1016/S0091-6749(97)70147-7) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 4 Eosinophil supernatants derived from subjects with IA or AD, but not from NA subjects, significantly prolong the survival of eosinophils from NA subjects in culture. Eosinophils from NA subjects were incubated for 72 hours with 300 μl medium or with 100 μl or 300 μl of eosinophil supernatants derived from subjects with AD (SUP-AD), IA (SUP-IA), or NA subjects (SUP-NA) after 72 hours in culture. Journal of Allergy and Clinical Immunology 1997 100, 536-543DOI: (10.1016/S0091-6749(97)70147-7) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 5 Neutralizing effect of anti–GM-CSF mAbs. Eosinophils from AD patients were incubated for 72 hours without (unfilled bars) or with (filled bars) anti–GM-CSF mAbs neutralizing 1000 U of GM-CSF bioactivity. Journal of Allergy and Clinical Immunology 1997 100, 536-543DOI: (10.1016/S0091-6749(97)70147-7) Copyright © 1997 Mosby, Inc. Terms and Conditions