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Adoptively transferred late allergic response is inhibited by IL-4, but not IL-5, antisense oligonucleotide  Sophie Molet, PhD, David Ramos-Barbón, MD,

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Presentation on theme: "Adoptively transferred late allergic response is inhibited by IL-4, but not IL-5, antisense oligonucleotide  Sophie Molet, PhD, David Ramos-Barbón, MD,"— Presentation transcript:

1 Adoptively transferred late allergic response is inhibited by IL-4, but not IL-5, antisense oligonucleotide  Sophie Molet, PhD, David Ramos-Barbón, MD, James G. Martin, MD, Qutayba Hamid, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 104, Issue 1, Pages (July 1999) DOI: /S (99) Copyright © 1999 Mosby, Inc. Terms and Conditions

2 Fig. 1 Schematic diagram representing one example of the mechanism by which AS ODNs work. AS ODNs act on the translation stage. Usually designed to hybridize specifically to a particular sequence of mRNA, AS ODNs block the access of mRNA to the ribosomes, thus preventing translation in the protein of interest. Then mRNA degradation is promoted by an RNase H-dependent agent (RNase H specifically degrades the RNA strand of RNA/DNA hybrid). Consequently, the production of a targeted protein is inhibited. Journal of Allergy and Clinical Immunology  , DOI: ( /S (99) ) Copyright © 1999 Mosby, Inc. Terms and Conditions

3 Fig. 2 Time course of R L after OVA challenge in BN rats. Unsensitized animals were injected intraperitoneally with CD4 + T cells treated with culture medium (controls [Co] , n = 9, open circles ) or NP-Y AS (irrelevant AS, filled circles , n = 13), IL-5 AS ( filled squares , n = 8), or IL-4 AS ( open triangles , n = 9) ODNs. Two days later, the animals were challenged with a 5% OVA aerosol. R L was measured every 5 minutes during a 20-minute period and subsequently every 15 minutes over a period of 8 hours. Journal of Allergy and Clinical Immunology  , DOI: ( /S (99) ) Copyright © 1999 Mosby, Inc. Terms and Conditions

4 Fig. 3 Airway response to OVA in BN rats. Unsensitized animals were injected intraperitoneally with CD4 + T cells treated with culture medium (controls [Co] , n = 9, hatched bar ) or NP-Y AS (irrelevant AS, open bar , n = 13), IL-5 AS ( shaded bar , n = 8), or IL-4 AS ( filled bar , n = 9) ODNs. Two days later, the animals were challenged with a 5% OVA aerosol. R L was measured for 8 hours and analyzed during the first hour for the development of an EAR (A) and from 1 to 3 hours (B) and 3 to 8 hours for the measurement of the LAR (C) . EAR was quantified by taking the maximal value of R L during the first 60 minutes after OVA challenge above the baseline (results expressed as percentage of baseline). LAR was calculated as the area under the R L versus time curve above the baseline values from 3 to 8 hours (results expressed as AUC). Results are expressed as means ± SEM. Probability (P) values were determined by using the nonparametric Kruskal-Wallis test. Journal of Allergy and Clinical Immunology  , DOI: ( /S (99) ) Copyright © 1999 Mosby, Inc. Terms and Conditions

5 Fig. 4 Representative photomicrographs of BAL cells recovered from control rats ( A and C ) and from rats that received IL-4 AS–treated CD4 + T cells ( B and D ). Specific staining shown is for MBP-positive eosinophils ( A and B ; black arrows ) and IL-4–immunoreactive cells ( C and D ; white arrows ). Journal of Allergy and Clinical Immunology  , DOI: ( /S (99) ) Copyright © 1999 Mosby, Inc. Terms and Conditions

6 Fig. 5 Eosinophilia in BAL fluid investigated by immunocytochemistry. BAL fluid was recovered at the end of the experiment (8 hours after OVA challenge) from the recipients of culture medium (controls [Co], n = 9, circles) or NP-Y AS– (irrelevant AS, triangles, n = 8), IL-5 AS– (squares, n = 8), or IL-4 AS– (diamonds, n = 8) treated CD4+ T cells. Eosinophils were detected by the APAAP method by using the BMK-13 antibody, a mouse anti-human MBP mAb. Results are expressed as the number of MBP+ cells per 103 BAL cells counted for each rat. Probability (P) values were determined by using ANOVA, and the Fisher test was applied to test for differences. Journal of Allergy and Clinical Immunology  , DOI: ( /S (99) ) Copyright © 1999 Mosby, Inc. Terms and Conditions

7 Fig. 6 IL-4 expression in BAL fluid investigated by immunocytochemistry. BAL fluid was recovered at the end of the experiment (8 hours after OVA challenge) from the recipients of culture medium (controls [Co], n = 9, circles) or NP-Y AS– (irrelevant AS, triangles, n = 8), IL-5 AS– (squares, n = 8) or IL-4 AS– (diamonds, n = 8) treated CD4+ T cells. IL-4 was detected by the streptavidin/biotin–alkaline phosphatase conjugated method by using a biotinylated goat anti-mouse IL-4 antibody. Results are expressed as the number of IL-4+ cells per 103 BAL cells counted for each rat. Probability (P) values were determined by using ANOVA, and the Fisher test was applied to test for differences. Journal of Allergy and Clinical Immunology  , DOI: ( /S (99) ) Copyright © 1999 Mosby, Inc. Terms and Conditions

8 Fig. 7 IL-5 expression in BAL fluid investigated by immunocytochemistry. BAL fluid was recovered at the end of the experiment (8 hours after OVA challenge) from the recipients of culture medium (controls [Co] , n = 9, circles ) or NP-Y AS– (irrelevant AS, triangles , n = 7), IL-5 AS– ( squares , n = 8) or IL-4 AS– ( diamonds , n = 8) treated CD4 + T cells. IL-5 was detected by the APAAP method by using a mouse anti-rat IL-5 mAb. Results are expressed as the number of IL-5 + cells per 10 3 BAL cells counted for each rat. Probability (P) values were determined by using ANOVA, and the Fisher test was applied to test for differences. Journal of Allergy and Clinical Immunology  , DOI: ( /S (99) ) Copyright © 1999 Mosby, Inc. Terms and Conditions

9 Fig. 8 IFN-γ expression in BAL fluid investigated by immunocytochemistry. BAL fluid was recovered at the end of the experiment (8 hours after OVA challenge) from the recipients of culture medium (controls [Co], n = 9, circles) or NP-Y AS– (irrelevant AS, triangles, n = 7), IL-5 AS– (squares, n = 8), or IL-4 AS– (diamonds, n = 8) treated CD4+ T cells. IFN-γ was detected by the APAAP method by using a mouse anti-rat IFN-γ mAb. Results are expressed as the number of IFN-γ+ cells per 103 BAL cells counted for each rat. Probability (P) values were determined by using ANOVA, and the Fisher test was applied to test for differences. Journal of Allergy and Clinical Immunology  , DOI: ( /S (99) ) Copyright © 1999 Mosby, Inc. Terms and Conditions

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