Figure 1.
Structural analysis of the tyrosine residues in Bet v 1.0101.
(A) Cartoon representation with alpha-helices in magenta and beta-strands in yellow. The tyrosine residues are shown as sticks. (B) Electrostatic potential mapped to the protein surface. Positive charges are colored blue and negative charges are colored red. The tyrosine residues are show as spheres, the CH groups are show in green color. (C) The front half of Bet v 1 is cut off providing a view to the large central cavity. Tyrosines Y81 and Y83 are exposed to the cavity. The figure was prepared with UCSF Chimera [34].
Figure 2.
Presentation of allergen derived naturally processed HLA-DR associated peptides by DCs from ten different donors (B01–B10).
Allergen derived peptides were detected in samples generated from unmodified Bet v 1 loaded DCs (light grey bars) and in samples generated from Bet v 1 nitro loaded DCs (dark grey bars). In the sequence, tyrosine residues are highlighted grey.
Table 1.
Allergen derived peptide clusters and peptide length variants identified in donors B01–B10.
Figure 3.
Nitration of Bet v 1 increases the copy number of identified allergen-derived peptides with identical amino acid sequence.
The numbers of detected copies of allergen derived peptides with identical peptide sequence, derived from DCs loaded with unmodified Bet v 1 (grey bars) and allergen derived peptides derived from DCs loaded with Bet v 1 nitro (black bars) are shown for each identical peptide and corresponding donors. Due to the limited amount of available cells for peptide isolation, each sample was measured only once.
Figure 4.
Bet v 1 specific T cell lines proliferate stronger in response to Bet v 1 nitro compared to unmodified Bet v 1 at 1.25 µg/ml (p = 0.046).
Each symbol indicates the results of a single donor (n = 6) and the median is indicated with a horizontal bar. Treatment with 1.25 µg/ml unmodified Bet v 1 or Bet v 1 nitro lead to a median proliferation value of 33293.5 delta cpm or 54157.0 delta cpm, respectively. Median proliferation for treatment with 1.25 µg/ml HSA or HSA nitro could be determined as 336.3 delta cpm and 840.8 delta cpm respectively.
Table 2.
HLA-DRB-alleles of the donors B01 to B10.