Figure 2.
Figure 2. CD8+ T cells (106) were stimulated with pp65 CMV peptide-pulsed T2 cells for 3 hours and then diluted logwise into unstimulated, autologous CD8+ T cells. The number of CMV-specific CD8+ T cells in the starting material determined by tetramer assay was used to calibrate CMV-specific CD8+ T cells in each dilution and correlate this with the number of IFN-γ mRNA copies such that the lowest concentration contained one CMV-positive CD8+ T cell/106 nonstimulated CD8+ T cells. RNA was then extracted for qRT-PCR. The lower limit of detection by the tetramer and PCR assay was 1/10 000 and 1/100 000 CMV-specific CD8+ T cells, respectively.

CD8+ T cells (106) were stimulated with pp65 CMV peptide-pulsed T2 cells for 3 hours and then diluted logwise into unstimulated, autologous CD8+ T cells. The number of CMV-specific CD8+ T cells in the starting material determined by tetramer assay was used to calibrate CMV-specific CD8+ T cells in each dilution and correlate this with the number of IFN-γ mRNA copies such that the lowest concentration contained one CMV-positive CD8+ T cell/106 nonstimulated CD8+ T cells. RNA was then extracted for qRT-PCR. The lower limit of detection by the tetramer and PCR assay was 1/10 000 and 1/100 000 CMV-specific CD8+ T cells, respectively.

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