Fig. 3

Enrichment of JC-targeted T cells. Top: After stimulation with five non-overlapping JC virus peptide libraries (after stimulation, before selection: ORI), cells were stained with anti-CD45 (FITC), anti-CD3 (APC), anti-CD4 (APC/Cy7), anti-CD8 (PACBlue) and anti-IFNγ (PE), washed and resuspended in acquisition buffer containing 7-AAD. Note a minimal frequency of IFNγ + CD4 + and CD8 + T-cells, yet the absence of IFNγ-bright T-cells. Bottom: Final product (TARGET), after immunomagnetic selection of ORI with anti-IFNγ immunomagnetic beads. JC virus + T-cells are recognized as 7AAD-/CD45bright/SSClow/CD3 + cells with IFNγ on the surface. Enrichment of JC-virus specific T-cells to 65% among CD4 + and 68% among CD8 + T-cells is observed