Fig. 4.

Selection of GP-100 (left panel) and NA17-A (right panel) specific T cells with M450-AvT multimers. a HLA-A*0201/GP100 or NA17-A monomers coated on M450-AvT were used to sort specific T cells from peptide stimulated PBMC from a melanoma patient. After in vitro ployclonal amplification, percentages of specific T cells were assessed by double staining with PE-conjugated tetramer and FITC-conjugated anti-CD8 antibody. b Repertoire diversity of sorted populations was assessed by labeling with 24 anti-Vb antibodies. c Avidities of M450-AvT sorted T cells for GP100 or NA17-A peptide were evaluated on peptide-loaded T2 cells as in Fig. 3. d Lysis of melanoma cell lines expressing both HLA-A2 and the relevant antigen [M113 (filled triangle) and M88 (open square) for gp100 sort and M17 (open circle) and DAGI (black square) for NA17-A] along with negative controls [M6 (open triangle) for HLA-A2 and M113 (filled triangle) for NA17-A] was measured at three differents E:T ratio (2:1, 10:1 and 50:1) by a 4 h ⁵¹Cr release assay. e Cytokine production of sorted cells in response to melanoma cell-lines was evaluated as in Fig. 3 [INFγ (grey square), IL2 (open square), TNFα (black square)]