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IL-4 upregulates FcϵRI α-chain messenger RNA in eosinophils

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Presentation on theme: "IL-4 upregulates FcϵRI α-chain messenger RNA in eosinophils"— Presentation transcript:

1 IL-4 upregulates FcϵRI α-chain messenger RNA in eosinophils
Nobuhisa Terada, MDa, Akiyoshi Konno, MDa, Yoshie Terada, MDa, Setsuya Fukuda, PhDb, Tetsuji Yamashita, PhDb, Tatsuya Abe, PhDc, Hiroko Shimada, PhDc, Kazuto Ishida, PhDc, Kentaro Yoshimura, PhDc, Youichi Tanaka, MDd, Chisei Ra, MD, PhDe, Kazuo Ishikawa, MDf, Kiyoshi Togawa, MDf  Journal of Allergy and Clinical Immunology  Volume 96, Issue 6, Pages (December 1995) DOI: /S (95) Copyright © 1995 Mosby, Inc. Terms and Conditions

2 FIG. 1 Expression of FcϵRIα mRNA in peripheral blood eosinophils of patients with nasal allergy and normal control subjects. See Table I for clinical details. PCR-assisted Southern blot analysis was performed as described in the text. Total RNA was isolated from fresh peripheral blood eosinophils. Sterilized water was used as a negative control. The human basophil cell line KU-812 was used as a positive control. Amounts of β-actin were adjusted to be almost equal between samples. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

3 FIG. 2 Expression of FcϵRIα mRNA and histamine H2 receptor mRNA in eosinophil-rich samples. PCR-assisted Southern blot analysis was performed as described in the text. Note that both FcϵRIα mRNA and histamine H2 receptor mRNA are observed in the human basophil cell KU-812. Peripheral blood eosinophils, however, expressed FcϵRIα mRNA but not histamine H2 receptor mRNA. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

4 FIG. 3 Effect of cytokines on the expression of FcϵRIα mRNA in peripheral blood eosinophils. Representative experiments. (A) Case 1 (nasal allergy). (B) Case 2 (nasal allergy). Total RNA was isolated from peripheral blood eosinophils after incubation for 6 hours with various concentrations of recombinant human IL-5, IL-4, IL-3, and and GM-CSF. PCR-assisted Southern blot analysis was performed as described in the text. Sterilized water was used as a negative control. The human basophil cell line KU-812 was used as a positive control. Amounts of β-actin were adjusted to be almost equal between samples. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

5 FIG. 3 Effect of cytokines on the expression of FcϵRIα mRNA in peripheral blood eosinophils. Representative experiments. (A) Case 1 (nasal allergy). (B) Case 2 (nasal allergy). Total RNA was isolated from peripheral blood eosinophils after incubation for 6 hours with various concentrations of recombinant human IL-5, IL-4, IL-3, and and GM-CSF. PCR-assisted Southern blot analysis was performed as described in the text. Sterilized water was used as a negative control. The human basophil cell line KU-812 was used as a positive control. Amounts of β-actin were adjusted to be almost equal between samples. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

6 FIG. 4 Effect of anti-monoclonal IL-4 antibody on the IL-4-induced enhancement in FcϵRIα mRNA levels. Eosinophils were incubated for 6 hours with 0.1 ng/ml recombinant human IL-4 in the presence or absence of various concentrations of anti-monoclonal IL-4 antibody. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

7 FIG. 5 Immunostaining pattern of FcϵRIα-positive eosinophils purified from peripheral blood. Some eosinophils were FcϵRIα-positive, as shown in A. Almost all eosinophils were FcϵRIα-negative (B). The FcϵRI+ human basophil cell line KU812 (C) served as positive control. Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

8 FIG. 6 Demonstration of FcϵRIα-positive cells in cryostat sections of nasal mucosa. Many FcϵRIα-positive cells are visible from the epithelial layer to the deep layer (A). Some FcϵRIα-positive cells exhibited typical eosinophil structure with polylobed nuclei (C, arrows). No FcϵRIα-positive cells were observed when sections were incubated with isotype-matched control antibody (B). Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

9 FIG. 6 Demonstration of FcϵRIα-positive cells in cryostat sections of nasal mucosa. Many FcϵRIα-positive cells are visible from the epithelial layer to the deep layer (A). Some FcϵRIα-positive cells exhibited typical eosinophil structure with polylobed nuclei (C, arrows). No FcϵRIα-positive cells were observed when sections were incubated with isotype-matched control antibody (B). Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

10 FIG. 7 Demonstration of FcϵRIα and ECP double-positive cells in cryostat sections of nasal mucosa. Double-labeling immunostaining for FcϵRIα and ECP-positive cells was peerformed as described in the text. Note that some ECP-positive cells are FcϵRIα-positive (arrows). Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

11 FIG. 7 Demonstration of FcϵRIα and ECP double-positive cells in cryostat sections of nasal mucosa. Double-labeling immunostaining for FcϵRIα and ECP-positive cells was peerformed as described in the text. Note that some ECP-positive cells are FcϵRIα-positive (arrows). Journal of Allergy and Clinical Immunology  , DOI: ( /S (95) ) Copyright © 1995 Mosby, Inc. Terms and Conditions

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