Fig. 4

Marker expression and cytokine release after restimulation with antigen. Functionality of expanded CMV-CTL obtained from G-CSF mobilized (n = 3) and non-mobilized (n = 3) PBMCs was analyzed by CD137 and CD69 expression quantification and cytokine production quantification after re-stimulation with CMVpp65-loaded PBMC (CMV feeders; CMV). (a) Mean CD137 and CD69 expression after CMV-loaded feeder re-stimulation, quantified from the CD3+ gate. (b) Surface marker expression in a representative re-challenge experiment for both non-mobilized and G-CSF mobilized samples. (c) Intracellular cytokine staining (ICS) method was used to determine the percentage of CD3+ cells that produced IFN-γ, IL-2, IL-10, TNF-α, and granzyme B (Granz B, Granzyme B). (d) Representative histograms of an ICS experiment for the IFN-γ, IL-2, IL-10, TNF-α, and granzyme B production analysis of G-CSF mobilized and non-mobilized CMV-CTLs, with filled peaks showing matched isotype controls