Molecular and functional analysis of the antigen receptor of Art v 1-specific helper T lymphocytes.
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 immunodominant T-cell epitope Art v 1(25-36) have an HLA-DRB1*01 phenotype.


OBJECTIVE
We studied cloning and functional expression of a human alphabeta 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 alpha-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 Vbeta18-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.


CONCLUSIONS
For the first time, a detailed molecular and functional analysis of a human allergen-specific TCR is presented.
Institute of Immunology, Center for Physiology, Pathophysiology and Immunology, Medical University of Vienna, Vienna, Austria.
Victoria M
VM
Leb
Beatrice
B
Jahn-Schmid
Klaus G
KG
Schmetterer
Hans J
HJ
Kueng
Daniela
D
Haiderer
Alina
A
Neunkirchner
Gottfried F
GF
Fischer
Karl
K
Nissler
Arnulf
A
Hartl
Josef
J
Thalhamer
Barbara
B
Bohle
Brian
B
Seed
Winfried F
WF
Pickl
D000485:Allergens / Q000276:immunology / Q000378:metabolism*; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D001402:B-Lymphocytes; D001483:Base Sequence; D002460:Cell Line; D002461:Cell Line, Transformed; D003001:Cloning, Molecular; D015234:HLA-A Antigens / Q000378:metabolism; D059811:HLA-DRB1 Chains; D006801:Humans; D019169:Jurkat Cells; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000378:metabolism*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013601:T-Lymphocytes; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology / Q000378:metabolism; D014161:Transduction, Genetic; D019076:Transgenes
D000485:Allergens; D052179:Antigens, Plant; C102724:Art v 1 protein, mugwort; D015234:HLA-A Antigens; D059811:HLA-DRB1 Chains; C487961:HLA-DRB1*01:01 antigen; D010940:Plant Proteins; D016693:Receptors, Antigen, T-Cell, alpha-beta