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Figure 2 | Journal of Translational Medicine

Figure 2

From: Whole blood assessment of antigen specific cellular immune response by real time quantitative PCR: a versatile monitoring and discovery tool

Figure 2

Cytokine gene expression induced by HCMV, EBV and influenza virus derived HLA class I restricted antigenic peptides in WB of healthy donors. WB from two HLA-A0201+ healthy donors (panels A and B), seropositive for HCMV and EBV (300 μl) was incubated for 5 hours in the presence of HCMV pp65495–503 (triangles), EBV LMP-2426–434 (diamonds), BMLF-1259–267 (squares) and IM58–66 virus (crosses) derived peptides at 10 μg/ml final concentration. Melanocyte derived GP100280–288 peptide (circles) was used as negative control. RNAlater was then added and total cellular RNA was purified, reverse transcribed and amplified in the presence of primers and probes specific for IFN-γ (full symbols) or IL-2 (empty symbols). Specific gene expression was analyzed by using, as reference, the expression of β-actin house keeping gene (y axes). A threshold of 2-fold increase in specific gene expression over control values was used as cut-off (dashed lines for IFN-γ and dotted lines for IL-2 gene expression, respectively). WB specimens of the same size (300 μl) from the same donors were simultaneously stained with the corresponding multimers and the number of antigen specific T cells (x axes) was evaluated and correlated with antigen driven gene expression data.

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