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. 2000 Jan 4;97(1):400–405. doi: 10.1073/pnas.97.1.400

Figure 2.

Figure 2

Recognition of the Melan-A/MART-151–73 peptide and UPCI-MEL 136.1 cell line by CD4+T cells of an HLA-DRB1*0401 normal donor. CD4+ T cells from an HLA-DRB1*0401 donor underwent three rounds of in vitro stimulation with autologous DC pulsed with the Melan-A/MART-151–73 peptide as described in Materials and Methods. Ten thousand of the resulting responder CD4+ T cells were incubated in a 20-h IFN-γ ELISPOT assay in the presence of T2.DR4 cells pulsed with the Melan-A/MART-151–73 peptide or gp100167–189 peptide (1 μg/ml), UPCI-MEL 136.1 cells +/− anti-HLA-DR antibodies (L243), or UPCI-MEL 136.1 cells +/− anti-HLA-A,B,C antibodies (W6/32). IFN-γ spots were developed and counted by computer-assisted video image analysis. Each bar represents the mean spot number of triplicates ± SD with 104 CD4+ T cells initially seeded per well. We show the data of one representative experiment of six performed. * indicate significant results, i.e., P < 0.05. The mean values of experimental and background counts were compared by using Student's unpaired t test.