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Fig. 9 | BMC Immunology

Fig. 9

From: An optimized IFN-γ ELISpot assay for the sensitive and standardized monitoring of CMV protein-reactive effector cells of cell-mediated immunity

Fig. 9

T-activated® pp65 and IE-1 CMV antigens stimulate a broad range of CMV-reactive effector cells. Comparative analysis of IFN-γ secreting cells by ELISpot (a) and flow cytometry (b) following stimulation of PBMC with T-activated® pp65 and IE-1 antigens. PBMC (4 replicates each) from six CMV-seropositive healthy donors (median age and range of 39 (22–55) years; 2 male and 4 female) were stimulated with T-activated® pp65 and IE-1 antigens according to the optimized IFN-γ ELISpot (a) and to the protocol of intracellular and surface marker staining and flow cytometry described in the Methods section (b). Bar graphs in (a) depict IFN-γ-dependent SFC per 200,000 PBMC, as before. Bar graphs in (b) represent the number of IFN-γ-expressing CD3+CD4+ (Th), CD3+CD8+ (CTL), CD3CD56+ (NK) and CD3+CD56+ (NKT-like) cells per 200,000 lymphocytes. Note that the Y-axis scales in (b) were adjusted for a better resolution of the respective data. Individual age and gender of donors were as follows: d120, 37-year-old male; d172, 55-year-old female; d290, 46-year-old female; d300, 41-year-old female; d343, 24-year-old female; d361, 22-year-old male

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