Forschungsdatenbank PMU-SQQUID

PMU-Autor/inn/en

Abstract

Background: Ninety-five percent of patients with mugwort allergy are sensitized to Art v 1, the sole major allergen in mugwort (Artemisia vulgaris) pollen. Sixty-nine percent of patients recognizing the single inummodominant T-cell epitope Art v 1(25-36) have an HLA-DRB1*01 phenotype. Objective: We studied cloning and functional expression of a human alpha beta T-cell receptor (TCR) specific for Art v 1(25-36). Methods: TCR chains were RT-PCR amplified from an Art v 1(25-36)-Specific T-cell clone, retrovirally transferred, and functionally tested in Jurkat T cells or alternatively in peripheral blood T lymphocytes of nonallergic individuals. Results: The et-chain of the TCR is composed of TRAV17 and TRAJ45 segments, and the beta-chain uses TRBV18, TRBD1, and TRBJ2-7. Analyses of 23 other Art v 1-specific T-cell clones did not reveal preferential usage of the TRAV17, TRBV18, or other TCR gene families. Efficient TCR transfer into Jurkat T cells was shown by binding of TCR V beta 18-specific mAb and DRB1*0101/Art v 1 tetramers. Transgenic Jurkat T cells specifically recognized syngeneic EBV B cells pulsed with Art v 1(25-36) peptide and artificial antigen-presenting cells expressing invariant chain::Art v 1 fusion proteins. Moreover, transfer of the TCR into peripheral blood lymphocytes generated T cells that were Art v 1 reactive. Activation of transgenic T cells by artificial antigen-presenting cells was strictly dependent on costimulation. Conclusion: For the first time, a detailed molecular and functional analysis of a human allergen-specific TCR is presented.

Useful keywords (using NLM MeSH Indexing)

Allergens/immunology

Allergens/metabolism*

Amino Acid Sequence

Antigens, Plant

B-Lymphocytes

Base Sequence

Cell Line

Cell Line, Transformed

Cloning, Molecular

HLA-A Antigens/metabolism

HLA-DRB1 Chains

Humans

Jurkat Cells

Molecular Sequence Data

Plant Proteins/metabolism*

Receptors, Antigen, T-Cell, alpha-beta*/chemistry

Receptors, Antigen, T-Cell, alpha-beta*/genetics

Receptors, Antigen, T-Cell, alpha-beta*/metabolism

T-Lymphocytes

T-Lymphocytes, Helper-Inducer/immunology

T-Lymphocytes, Helper-Inducer/metabolism

Transduction, Genetic

Transgenes

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