Table 1

Staining of Melan-A/Mart-1-specific T cells by HLA-A*0201-Melan-A/Mart-126–35 tetrameric complexes

Staining with HLA-A*0201-Melan-A/Mart-126–35 PE-tetramers or HLA-A*0201-influenza-matrix58–66 PE-tetramers was performed on: PBLs of a HLA-A*0201 donor (negative control); a Melan-A/Mart-1-specific CTL clone A83 (25) , an influenza-matrix58–66-specific T cell line (25) , fresh TILs from a melanoma metastasis previously shown to contain a high frequency of Melan-A/Mart-1-specific T cells (22) (positive controls); and PBLs of a HLA-A*0201+ patient (Pt 1) enrolled in the rHuIL-12 study. Tetramer staining with positive and negative controls was used to set markers on FL-2 histograms to evaluate the numbers of tetramer+ T cells in the experimental samples from patient (Pt) 1. Frequency of tetramer+ T cells obtained by single staining with tetramers was confirmed by two-color analysis on cells stained with PE-labeled tetramers and FITC-labeled anti-CD8 mAbs (data not shown). PBLs of patient 1 were taken before (day −4) and after (day +1, +4, and +7) the first rHuIL-12 injection. One × 106 cells were analyzed for each sample.

Lymphocytes fromExperiment no.Tetramer cells/106 cells after staining with:
HLA-A*0201 donor00
Influenza-matrix58–66-specific T cell line0450,669
CTL A83977,5000
Fresh TILs57,3401,937
PBLs of Pt 1, Day−410775
PBLs of Pt 1, Day+11252208
PBLs of Pt 1, Day +41239120
PBLs of Pt 1, Day+42255161
PBLs of Pt 1, Day+7101,172
PBLs of Pt 1, Day +7201,048