Table 1.

Effects of isothermal dose with temperature below the breakpoint temperature (41.8°C/120 min) on 624.38-MEL

	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