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. Author manuscript; available in PMC: 2016 Jul 1.
Published in final edited form as: Clin Cancer Res. 2015 Feb 17;21(13):2952–2962. doi: 10.1158/1078-0432.CCR-14-2998

Figure 1. K562-based whole-cell vaccine encoding CMV-pp65 and CD40L matures monocytes and stimulates CMV-CTLs in vitro. Panel A.

Figure 1

Expression of CD40L and OX40L in engineered K562. Striped histograms indicate wild type K562 cells. Panel B. Western blot showing the expression of CMV-pp65 in engineered K562. Panel C. Uptake of apoptotic bodies from irradiated K/pp65 and K/CD40L/pp65 by monocytes. Monocytes labeled with PKH26 red fluorescent cell linker compound were co-cultured (5:1 ratio) with irradiated K/CD40L/pp65 labeled with PKH2 green fluorescent cell linker compound. Analysis of fluorescence signals was performed after 72 hours of co-culture using a fluorescence microscope (Olympus IX70). Panel D. Expression of CD80, CD83, CD11c and HLA-DR by monocytes 72 hours after co-culture with irradiated K/pp65 (in blue) and K/CD40L/pp65 (in green). The red line represents the expression of CD80, CD83, CD11c and HLA-DR before the stimulation. Panel E. Frequency of CMV-CTLs assessed by IFNγ ELISpot using the CMV-pp65 pepmix. Data represented mean ± SD of 11 CMV-seropositive donors. Stimulation with an irrelevant pepmix was used as a negative control.