Location and modification of potential target peptides within PASD1.

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Location and modification of potential target peptides within PASD1. Location and modification of potential target peptides within PASD1. (A) Location of wild peptides (a.a. numbers shown) on variant 1 (_v1) and variant 2 (_v2) of PASD1. GKT-ATA20 is the region of PASD1 identified by AML patient sera (8). (B) Stabilization of HLA-A2 molecules on the surface of T2 cells using wt (Pw8) or analogue peptides (Pa13 and Pa14). A peptide from CMV was used as a positive control. Faint line: FITC-isotype control; bold line: FITC-HLA-A2-specific antibody. (C) Duration of binding of peptides to T2 cells was measured at various time-points after the removal of peptide by FACS analysis of HLA-A2 levels. (D) Purified CD3+ T cells were stimulated with autologous DCs loaded with peptides Pa11–Pa16, CMV, FLU, or no peptide. The positive control (Flu or CMV), which gave optimal results, is shown. Aliquots of culture supernatant were collected on day 7 and analyzed for IFN-γ production levels by ELISA. Nicola Hardwick et al. Cancer Immun 2013;13:16 Copyright © 2013 by Barbara-ann Guinn