Table 1.
HLA-A2-peptide induced direct ex vivo IFN-γ ELISPOT responses of HLA-A2 positive healthy donors' PBMCa
| Donors: | #1 | #2 | #3 | #4 | #5 | #6 |
|---|---|---|---|---|---|---|
| HLA-A allele “a” | 2-0201 | 2-0201 | 2-0201 | 2-0201 | 2-0201 | 2-0201 |
| HLA-A allele “b” | 33-3301 | 26-2601 | 2-0201 | 29-2902 | 24-2402 | 3-0301 |
| HLA-A2-restricted peptides: | ||||||
| Flu Mat 158 (GILGFVFTL) | 4 +/- 1 | 40+/- 12 | 72 +/- 38 | 28 +/- 4 | 33 +/- 1 | 4 +/- 0 |
| Flu A (FMYSDFHFI) | 24 +/- 10 | 37+/- 15 | 594 +/-142 | 32 +/- 6 | 32 +/- 3 | 6 +/- 0 |
| EBV LMP2A (CLGGLLTMV) | 139 +/- 15 | 3 +/- 1 | 779 +/-106 | 2 +/- 1 | 1 +/- 1 | 1 +/- 0 |
| EBV BMLF1259 (GLCTLVAML) | 13 +/- 1 | 12+/- 6 | 17 +/-3 | 7 +/- 1 | 4 +/- 1 | 35 +/- 11 |
| HCMV pp65 495 (NLVPMVATV) | 126 +/- 2 | 3 +/-1 | 762 +/-144 | 2 +/- 0 | 3 +/- 1 | 1 +/- 0 |
| CEF peptide pool (32 peptides) | 232 +/- 21 | 81 +/- 29 | 796 +/-60 | 273 +/-47 | 75 +/- 4 | 71 +/- 18 |
PBMC (2 × 105/ well) from the specified HLA-A2 positive donors (identified by high resolution typing) were cultured with the specified HLA-A2- or non-HLA-A2- restricted peptides (sequence specified, tested at 2 µg/ml) for 24 hr and the peptide-induced IFN-γ production was measured in a standard IFN-γ ELISPOT assay. CEF peptide pool (containing 32 peptides of EBV, CMV and flu) was also tested. In the negative control wells spot counts were smaller than 3 spots per well. The data are shown as mean IFN-γ SFU per 200,000 PBMC ± SD for triplicate wells for one experiment that has been reproduced twice with similar results – the inter-assay variation of peptide-induced responses was < 30%.