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. Author manuscript; available in PMC: 2014 Jan 24.
Published in final edited form as: J Immunol. 2010 Dec 3;186(1):312–322. doi: 10.4049/jimmunol.1001989

FIGURE 3.

FIGURE 3

Peptides LAGE-197–114, LAGE-1104–121, and LAGE-1b118–135 represent promiscuous HLA-DR epitopes recognized by CD4+ T cells isolated from PBLs of melanoma patients. CD4+ T cells were isolated from PBLs of melanoma patients and restimulated four times on a weekly basis with irradiated autologous mature DCs pulsed with individual peptide LAGE-197–114 (A), LAGE-1104–121 (B), or LAGE-1b118–135 (C). CD4+ T cells (1 × 104) were incubated in 48-h IFN-γ ELISPOT assays in the presence of L.DR cells pulsed with relevant LAGE-1–derived peptide or irrelevant peptide LAGE-1 ORF285–102. Spots were developed and counted by computer-assisted video image analysis. Each bar represents the mean spot number of triplicates (±SD) with 1 × 104 CD4+ T cells initially seeded per well. *p < 0.05. One representative experiment of at least three performed is depicted.