Cross-stimulation of CD14- cells by mimic peptides.

<p>CD14⁻ cells were incubated with DCs loaded with mimic peptides and then re-stimulated with a fresh batch of DCs loaded with same or a different mimic peptide. T-cell stimulation was monitored by quantifying the amount of IFN-γ released into the medium. The IFN-γ levels were normalized to the levels produced following incubation with DCs that had no peptides loaded. Error bars represent the normalized % error from two independent experiments. The combination of peptides are indicated in the figure and are as follows <b>A.</b> MP1 followed by MP1; red solid line (•), MP1 followed by MP5; red dashed line (▪), MP5 followed by MP5; green solid line (▴) and MP5 followed by MP1; green dashed line (⋄). <b>B.</b> MP3 followed by MP3; black solid line (▴), MP4 followed by MP4; green solid line (•), MP4 followed by MP3; red solid line (▪).</p

Physical properties of recogniser peptides.

<p>Physical properties of recogniser peptides.</p

Physical properties of MUC1pep1, MUC1pep2 and their putative mimic peptides MP1-5.

<p>Physical properties of MUC1pep1, MUC1pep2 and their putative mimic peptides MP1-5.</p

MUC1 peptides and their corresponding recogniser peptides.

*<p>From <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0049728#pone.0049728-Luo1" target="_blank">[35]</a> Sequence similarities between anti-MUC1 CDRs and recognisers are shown in bold and underlined.</p

Immunostimulation of CD14- cells with peptide-loaded DCs.

<p><b>A.</b> CD14⁻ cells were co-cultured with DCs loaded with MUC1pep1 core peptide (dotted line) or MUC1pep2 core peptide (solid line). After 8 days (arrow), the cells were re-stimulated by addition of a fresh batch of autologous DCs. T-cell stimulation was monitored by quantifying the amount of IFN-γ released into the medium. The IFN-γ levels were normalized to the levels produced following incubation with DCs that had no peptides loaded. Error bars represent the normalized % error from two independent experiments. <b>B.</b> As in A except that DCs were loaded with MP1 (solid line), MP2 (dashed line) or MP3 (dotted line). <b>C.</b> As in A except that DCs were loaded with MP4 (dashed line) and MP5 (solid line).</p

MUC1 peptides, corresponding recognisers and putative mimic peptides.

<p>Sequence similarities between mimic peptides (MP2 and MP3) and MUC1pep1 are shown in bold and underlined.</p

Structure of the MUC1 protein.

<p>The secretory signal peptide is shown as a shaded box at the N-terminus. The variable number tandem repeat (VNTR) is shown as an open rectangle. The small C-terminal domain, which is proteolytically derived from a common precursor protein and containing the transmembrane domain is also shown. The expanded region shows the amino acid sequences of three VNTR repeats [in brackets]. The amino acid sequences of the 12-mer and 14-mer peptides (MUC1pep1 and MUC1pep2 respectively) used for biopanning are underlined. The sequence of the core peptides used for eluting bound phage, competition ELISA and DC stimulation are shown in red.</p