Table 1.
Control | Recovery at 37°C | |||||
---|---|---|---|---|---|---|
37°C | 4 h | 15 h | 24 h | 48 h | 72 h | |
HLA-A2ΔMFI a | 458±92 | 354±131 | 437±157 | 511±165 | 584±169 | 669±59 |
Hsp70 ΔMFIa | 47±27 | 198±50 | 265±56 | 383±67 | 419±83 | 385±79 |
Tyosinaseb | 1 | 0.80±0.12 | 0.76±0.06 | 0.96±0.12 | 1.32±0.06 | Nt |
IFN-γ of TyrF8c | 100 | 71±10 | 67±9 | 88±16 | 85±8 | 103±13 |
IFN-γ of A42c | 100 | 79±10 | 86±9 | 89±11 | 90±6 | 97±6 |
aΔMean fluorescence intensities (MFI) were calculated by subtracting the MFI value of the isotype control (MOPC21) from the MFI value of the specific antibody HB54 (directed against HLA-A2 molecules) or 6B3 (directed against inducible HSp70 molecules). MFI of isotype control ranged between 3 and 25. Results are the mean values of ΔMFI ± SD from four to three independent experiments.
bValues are the fold-change in transcript levels relative to the level at 37°C, which was set to one (crossing points at 37°C were approximately 16 for tyrosinase). Values are the mean of 3 independent experiments (± SD). The confidence interval in which all values show a difference of ±1.5 cycles compared to the reference value at 37°C discriminate between significant overexpression of transcripts from significant underexpersion (0.35–2.8)
cValues are the % of IFN-γ relative to control cells at 37°C, which was set to 100%. TyrF8 and A42 are two CTL clones that recognize tyrosinase-peptide and Melan-A/MART-1 peptides, respectively. IFN-gamma was measured in supernatants of 24 h-cocultures of CLT with treated 624.38-MEL