Table 3

Quantitation of the number of D6-HLA-A2 complexes on the surface of cells

CellsMean number of sites/cella
JY pulsed (1 μm peptide)120,959 ± 16,934
MCF-7 pulsed (1 μm peptide)5,539 ± 1,828
MDA-MB-231 (1 μm peptide)10,022 ± 2,004
MCF-7284 ± 170
MDA-MB-231173 ± 112
Backgroundb26 ± 11
  • a The fluorescence intensity of stained cells in each experiment was compared with fluorescence intensities of calibration beads with known numbers of PE molecules/bead (QuantiBRITE PE beads, Becton-Dickinson), and the number of sites for each experiment was determined. The number of nonspecific sites was determined by the intensity of staining of cells that do not express MUC1 or HLA-A2. The number of specific sites for each experiment was then calculated for each experiment. The mean number of sites was calculated from 10 independent experiments/determinations. The deviation in number of sites depend on the sensitivity of detection and the physiological status of the cells in each individual determination.

  • b Background number of sites was determined as described previously, using SK-BR3 (HLA-A2/MUC1+), FM3D (HLA-A2+/MUC1), and G-43 (HLA-A2/MUC1) cells as controls.