Anti tim 3 clone f38 2e2

Phenotypic characterization on resting T cell clones was performed on 10⁵ T cells labeled with MELOE-1 and Melan-A tetramers (10 μg/mL) (Recombinant protein facility, SFR Santé, Nantes, France), anti-CD8 (clone BW135/80, Miltenyi Biotec), anti-CD45RO (clone UCHL1, BD Biosciences), anti-CD27 (clone M-T271, BD Biosciences), anti-CD28 (clone CD28.2, BD Biosciences), anti-CD62L (clone DREG-56, BD Biosciences), anti-PD-1 (clone EH12, BD Biosciences), anti-CTLA-4 (clone BNI3, Miltenyi Biotec), anti-BTLA (clone J168–540, BD Biosciences), anti-Tim-3 (clone F38–2E2, eBiosciences) and anti-CD95 (clone DX2, BD Biosciences) specific antibodies. PD-1 expression (Clone EH12, BD Biosciences) was tested on specific T cell clones or sorted T cells at rest and after activation by quadruple labeling with specific tetramers, anti-CD8 and anti-CD25 (clone M-A251, BD Biosciences), as activation marker. All the antibodies were used at a concentration of 5 μg/mL. Vß diversity of sorted Melan-A-specific T cell lines was analyzed by labeling with 24 anti-Vß mAbs included in the IOTest Beta Mark TCR V Kit (Beckman-Coulter, IM3497). All the cytometric analyses were performed on a Facs Canto II (BD Biosciences).