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Preserved epitope‐specific T cell activation by recombinant Bet v 1‐MBP fusion proteins
Author(s) -
R. J. Joost van Neerven,
Sparholt,
Schou,
Martin R. Larsen
Publication year - 1998
Publication title -
clinical and experimental allergy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.462
H-Index - 154
eISSN - 1365-2222
pISSN - 0954-7894
DOI - 10.1046/j.1365-2222.1998.00259.x
Subject(s) - epitope , fusion protein , microbiology and biotechnology , maltose binding protein , t cell , recombinant dna , biology , allergen , immune system , biochemistry , gene , antigen , immunology , allergy
Background Allergen‐specific T cells play an important role in the allergic immune response, and are thought to be the principal target in specific immunotherapy. Objective The aim of the present study was to evaluate if fusion proteins of allergens with bacterial proteins can be used to activate and bias allergen‐specific T cells, and to characterize T cell epitopes. Methods The complete gene of Bet v 1, the major birch pollen allergen, was amplified by PCR from birch pollen mRNA, and cloned in pKK223–3. The complete gene or truncated sequences were transferred to pMAL‐c and expressed in E. coli as fusion proteins with maltose binding protein (MBP). The complete fusion protein, and the truncated fusion proteins were used for studies with Bet v 1‐specific T cells. Results Bet v 1‐specific T cells reacted similarly with purified and crude Bet v 1‐MBP proteins. Therefore, crude preparations were used to study the epitope‐specificity of 11 Bet v 1‐specific T cell clones. Six distinct T cell epitopes were determined in this way. Interestingly, the T cell epitope of three T cell clones, that did not react with synthetic peptides in a previous study, was identified. In addition, the presence of MBP as a fusion partner to Bet v 1 was shown to influence TH 2 /TH 1 cytokine production in T cell lines, but not in established T cell clones. Conclusion Using crude preparations of recombinant fusion proteins of Bet v 1 with MBP, multiple T cell epitopes were identified in Bet v 1. As T cell activation is preserved in this system, the generation of recombinant allergens with TH 1 ‐inducing proteins as fusion partners might be considered as a T‐cell targeted approach for specific immunotherapy.