Definition of an alternative Tp2 epitope that binds to BoLA-6*04101. Panel A: p-MHC class I monomer formation ELISA assay using Tp227-37 and Tp229-37 peptides. Panel B: ELISpot performed on two clones (5 and 7) of the cell line BW02 with cell media alone, Tp1214-224, Tp227-37 or Tp229-37 peptides. Data are presented as spot forming units (SFU) and each well contained 2.5 × 104 CTLs. Asterisk (*) indicates saturated wells. Panel C: Two clones of the cell line BW02 was stained with anti-bovine CD8 (PerCP) and different BoLA-6*04101 tetramers. The unstained control or stained with the unfolded (no peptide) BoLA-6*04101 tetramer are depicted on the left. Tetramer labelling was measured on the CD8-gated cell population.