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Catherine M. Counsell, MAa, Julian F. Bond, PhDa, John L

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Presentation on theme: "Catherine M. Counsell, MAa, Julian F. Bond, PhDa, John L"— Presentation transcript:

1 Definition of the human T-cell epitopes of Fel d 1, the major allergen of the domestic cat 
Catherine M. Counsell, MAa, Julian F. Bond, PhDa, John L. Ohman, MDb, Julia L. Greenstein, PhDa, Richard D. Garman, PhDa  Journal of Allergy and Clinical Immunology  Volume 98, Issue 5, Pages (November 1996) DOI: /S (96) Copyright © 1996 Mosby, Inc. Terms and Conditions

2 FIG. 1 Complete primary structure of Fel d 1 chain 1 and Fel d 1 chain 2 and overlapping Fel d 1 synthetic peptides used for T-cell epitope mapping. Peptides 2-13 and represent the known polymorphism (long and short forms, respectively) in chain 2 of Fel d 1. Asterisks indicate amino acid deletions in the short form of Fel d 1 relative to the long form.12 Fel-1 and Fel-2 have previously been referred to as IPC-1 and IPC-2, respectiviely, in a previous publication.24 Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

3 FIG. 2 Prolifierative responses of two Fel d 1-reactive T-cell lines to overlapping Fel d 1 synthetic peptides and whole Fel d 1 protein. Short-term polyclonal T-cell lines were derived from the PBLs of two cat-allergic subjects, 736 (A) and 951 (B), as described in Methods, and were assayed for proliferation in response to the antigens indicated with irradiated autologous PBLs as antigen-presenting cells. Each antigen was titrated (3, 15, or 75 μg/ml), and the highest level of proliferation, regardless of dose, is presented. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

4 FIG. 2 Prolifierative responses of two Fel d 1-reactive T-cell lines to overlapping Fel d 1 synthetic peptides and whole Fel d 1 protein. Short-term polyclonal T-cell lines were derived from the PBLs of two cat-allergic subjects, 736 (A) and 951 (B), as described in Methods, and were assayed for proliferation in response to the antigens indicated with irradiated autologous PBLs as antigen-presenting cells. Each antigen was titrated (3, 15, or 75 μg/ml), and the highest level of proliferation, regardless of dose, is presented. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

5 FIG. 3 T-cell reactivity profile of responses to Fel d 1 peptides by short-term polyclonal Fel d 1-reactive T-cell lines derived from 53 subjects allergic to cats. Three concentrations (3, 15, or 75 μg/ml) of synthetic Fel d 1 peptides were tested for ability to stimulate proliferation of 53 Fel d 1-reactive polyclonal T-cell lines. The highest Sl for each peptide for each T-cell line was noted, regardless of dose. Sls for responses to each peptide were summed and provide an estimation of relative reactivity of regions of the Fel d 1 molecule in this population of subjects allergic to cats. The average medium control response was 717 cpm. As an example of positive responses (Sl ≥2.0), the average response to Fel 1-3 was 5058 cpm. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

6 FIG. 4 A comparison of Fel d 1 protein, Fel d 1 peptide Fel-1, and Fel d 1 peptide Fel-2 in the activation of short-term polyclonal Fel d 1-reactive polyclonal T-cell lines. Fel d 1-reactive T-cell lines were established from 53 subjects allergic to cats, as described in Methods, and were tested for proliferation in response to three concentrations (3, 15 or 75) μg/ml) of synthetic Fel d 1 peptides. The highest Sl for each peptide for each T-cell line was noted, regardless of dose. Results are presented as Pl for each antigen response, which is the product of mean Sl and frequency of response of at least twofold over medium control. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

7 FIG. 5 IgE reactivity of Fel d 1 (solid bars), peptide Fel-1 (hatched bars), and peptide Fel-2 (open bars). A, Direct ELISA of specific IgE directed against Fel d 1, Fel-1, and Fel-2 in pooled plasma from 20 subjects allergic to cats. B, Release of histamine in response to Fel d 1, Fel-1, and Fel-2 from peripheral blood basophils of 10 individual subjects allergic to cats. These data represent the maximum release for each antigen over a range of 0.5 to 2000 nmol/L. O.D., Optical density. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

8 FIG. 5 IgE reactivity of Fel d 1 (solid bars), peptide Fel-1 (hatched bars), and peptide Fel-2 (open bars). A, Direct ELISA of specific IgE directed against Fel d 1, Fel-1, and Fel-2 in pooled plasma from 20 subjects allergic to cats. B, Release of histamine in response to Fel d 1, Fel-1, and Fel-2 from peripheral blood basophils of 10 individual subjects allergic to cats. These data represent the maximum release for each antigen over a range of 0.5 to 2000 nmol/L. O.D., Optical density. Journal of Allergy and Clinical Immunology  , DOI: ( /S (96) ) Copyright © 1996 Mosby, Inc. Terms and Conditions

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