α1(V) peptide binding to HLA-DR1 and HLA-DR15.

<p>ProImmune REVEAL MHC-peptide binding assay screen of 5-amino acid overlapping peptide library (101 total peptides) of the α1(V) helical domain to HLA-DR15 (A), and HLA-DR1 (B). Each MHC:peptide binding score is based on known binding of a positive control peptide and a weaker binding positive control peptide. MHC:peptide binding scores above 12% are considered positive and have potential for immunological activity. Lack of bar for a peptide represents a score of 0%. Peptide number corresponds to the location in the α1(V) AA sequence. Blue shaded areas indicate epitope regions where only HLA-DR15 bound, red shaded areas where only HLA-DR1 bound, and yellow shaded areas indicate both HLA-DR1 and –DR15 bound within that helical domain.</p

Candidate α1(V) peptide screen in col(V)-responsive patients.

<p>PBMC from (A) CAD patient 20 (HLA-DR15, 17; -DQ2,6), (B) End-stage lung disease patient L138 (HLA-DR13,15; -DQ5,6) (C) Heart transplant recipient H76 (HLA-DR1,4; -DQ3,5, with donor HLA-DR9,11; -DQ3,-), (D) CAD patient 23 (HLA-DR11,17; -DQ2,7) and (E) End-stage lung disease patient PPG25 (HLA-DR7,11; -DQ2,7) were screened for reactivity to ProImmune binding peptides by TV-DTH. Results from HYP and non-HYP p629, p599 were combined. Patient response to intact col(V) (black bar), HLA-DR1-specific peptide (light gray bar), HLA-DR1/15-specific peptides (hatched bars), HLA-DR15-specific peptide (dark gray bar), and HLA-DQ/H-2 I-A^b binding peptides (white bars) are shown. (Data represents the mean of 2-4 separate assays).</p

Candidate α1(V) peptide screen in col(V) immunized HLA-transgenic mice.

<p>Splenocytes and ILN cells from col(V) immunized HLA-A*0201/DRB1*0101 Tg (open bars) and HLA-DRB1*1501 Tg (hatched bars) mice were pooled and co-injected with col(V), col(I), or candidate α1(V) peptides into footpads of CB-17 SCID mice to determine peptide-specific swelling responses ; n=3-6 mice were used for each test. * p=.026; # p=.057 comparing peptide responses in DRB1*0101 vs. DRB1*1501 Tg mice using an unpaired Mann-Whitney U test. Responses to col(I) negative control and col(V) positive control were not significantly different between the two Tg mouse strains.</p

Unusual pattern of µ1(V) peptide response in a DR1⁺ lung recipient with a DR15⁺ transplant donor.

<p>PBMC from lung transplant recipient (L86, HLA-DR1,11; -DQ3,5, with donor: HLA-DR1,15; -DQ5,-) was assayed by TV-DTH for reactivity to α1(V) peptides or intact col(V). Results from HYP and prolinated p629, p599 were combined. Patient response to intact col(V) (black bar), HLA-DR1-specific peptide (light gray bar), HLA-DR1/15-specific peptides (hatched bars), HLA-DR15-specific peptide (dark gray bar), and HLA-DQ/H-2 I-A^b binding peptides (white bars) are shown. Data represents the mean of 2-5 independent experiments. </p