Figure 4.

Expression of effector mediators by vaccine-activated Melan-A–specific T cells. Multiparameter cytometric analysis and sorting was performed with A2/Melan-A multimers and antibodies specific for CD8, CD45RA, and CCR7. (A) Expression of CD45RA and CCR7 is shown for A2/Melan-A multimer⁺ CD8⁺ gated cells. (B) RT-PCR gene expression analysis was performed (33) using primers specific for CD3, granzyme B, perforin, IFN-γ, TNF-α, and CD94, on sorted A2/Melan-A multimer⁺ T cells which were RA⁺CCR7⁺ (naive cells) or RA^–CCR7⁺ effector memory cells. Each band represents the RT-PCR product from RNA isolated from sorted 5-cell aliquots. Data in A and B (representative of 8 and 2 patients analyzed, respectively) are from PBMCs collected from patient LAU 371 before and after 2 and 4 vaccinations. +, positive control; –, negative control; N, naive T cells; EM, effector memory T cells.