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Rachel L. Miller, MDa, Thomas M

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1 Analysis of cytokine signaling in patients with extrinsic asthma and hyperimmunoglobulin E 
Rachel L. Miller, MDa, Thomas M. Eppinger, MDb, David McConnell, MDa, Charlotte Cunningham-Rundles, MD, PhDc, Paul Rothman, MDd  Journal of Allergy and Clinical Immunology  Volume 102, Issue 3, Pages (September 1998) DOI: /S (98) Copyright © 1998 Mosby, Inc. Terms and Conditions

2 Fig. 1 IL-4, IL-13, and IFN-γ expression during IgE-mediated disease. Sera cytokine levels were measured by ELISA. Data are shown as mean ± SE. IL-4 levels were significantly greater in patients with HIE than nonatopic control subjects, whereas IL-13 and IFN-γ levels did not differ among patients with HIE, asthma, or nonatopic control subjects. **P = .005. Journal of Allergy and Clinical Immunology  , DOI: ( /S (98) ) Copyright © 1998 Mosby, Inc. Terms and Conditions

3 Fig. 2 Western blot of whole cell extracts derived from PBMCs of 3 different representative subjects with asthma and simultaneously recruited nonatopic control subjects probed with α-Stat6 (A) and α-β-actin (B). Ramos cells and PBMCs (nonatopic control subjects) were stimulated with recombinant human IL-4 (100 U/mL) for 15 minutes before generation of whole cell extracts. Although nonactivated Stat6 is present in all asthmatic subjects (A) and nonatopic control subjects (C) represented here and in all subjects not shown, levels are not increased in patients (A). β-Actin levels are equivalent in all specimens (B). Journal of Allergy and Clinical Immunology  , DOI: ( /S (98) ) Copyright © 1998 Mosby, Inc. Terms and Conditions

4 Fig. 3 EMSA of representative patient with HIE. Whole cell extracts of PBMCs were probed with gas element from IRF-1 gene. Ramos cells and PBMCs stimulated in vitro for 15 minutes with recombinant IL-4 (100 U/mL) served as positive controls for stat6. Stat6 activation is not detected in this patient with HIE and markedly elevated IgE (9268 IU/Ml) and IL-4 level (174.7 pg/mL) or in any other patients or nonatopic control subjects tested. Journal of Allergy and Clinical Immunology  , DOI: ( /S (98) ) Copyright © 1998 Mosby, Inc. Terms and Conditions

5 Fig. 4 Southern blot of DNA extracted from PBMCs isolated from 2 patients with asthma (A), 2 patients with HIE, and 1 nonatopic control subject (C). U266 (IgE-secreting myeloma cells) served as positive controls. DNA was amplified by using nested primer PCR of switch fragments (Sμ/Sϵ), and specific products were determined by Southern transfer, hybridization, and incubation with a probe against Sϵ. Switch fragments are detected in U266 cells, as well as in 2 of 8 patients with asthma (1 case shown in second lane) and 2 of 3 patients with HIE (1 case shown in sixth lane) but not in any of the nonatopic control subjects. The last lane represents no DNA (negative control). Journal of Allergy and Clinical Immunology  , DOI: ( /S (98) ) Copyright © 1998 Mosby, Inc. Terms and Conditions

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