4.2. Flow staining

TCR epitope specificity was additionally validated by activating Jurkat-NFAT-GFP-TCR cells. The stained cells were then analyzed by flow cytometry to confirm surface expression and binding specificity for the two methods:

Co-culturing with K562-HLA Peptide Expressing Cells (CMV, EBV, and FLU)

Fig 6. Jurkat cells and K562 cells (1 x 10^5 each) were co-cultured in 96-well plate for 20hr for CMV, FLU and EBV peptides. FACS results show activation of the jurkat reporter cells after incubation as compared to negative control (K562 cells with negative control peptide).

Peptide-pulsing with K562-HLA cells (CMV, MART1, P53)

Fig 7. K562_HLA_A cells were incubated in serum-free medium with a specific peptide for 90 min. After washing once with complete medium, peptide-pulsed K562_HLA_A cells were used for coculture with Jurkat-NFAT-GFP reporter cells expressing specific TCR.