Flow cytometry immunophenotyping
Human antibodies specific for CD3 (SK7, Biolegend), CD4 (SK3,
Biolegend), CD8 (RPA-T8, BD Biosciences), CD45RA (HI100, Biolegend),
CD45RO (UCHL1, Biolegend), CCR7 (REA108, Miltenyi Biotec), CD95 (DX2 ,
Biolegend) were used for low cytometry acquisition for the determination
of Tn, Tcm, Tem, Tte phenotype. 8 color immunophenotyping kit specific
for CD3, CD5/16, CD19, CD14, CD45 (Miltenyi Biotec) were used for flow
cytometry acquisition for the determination of T cell purity, B cell, NK
cells, NKT cells, and monocytes percentage. MAGE-B2 TCR dextramer
(Immudex) and anti-human TCR Vβ13.2 (H132, Biolegend) were used to
detect MAGE-B2 TCR expression on CD8+ T cells and CD4 + T cells. FACS
Symphony or FACS Fortessa (BD Biosciences) was used for flow cytometry
acquisition. Samples were analyzed with FlowJo software.