In vitro CTL responses in HHD mice immunized with recombinant lv-hTERT vector
Peptides . | Sequence . | Relative avidity . | Protein of origin . | CTL response . | |
---|---|---|---|---|---|
R/T . | Percentage specific lysis . | ||||
p152 | LLARCALPV | Low | hTERT | 0/6 | — |
pY152 | YLARCALPV | High | hTERT | 0/6 | — |
p540 | ILAKFLHWL | High | hTERT | 5/6 | 64, 48, 57, 65, 41 |
p555 | ELLRSFFYV | Low | hTERT | 0/6 | — |
pY555 | YLLRSFFYV | High | hTERT | 1/6 | 16 |
p572 | RLFFYRKSV | Low | hTERT/mTERT | 5/6 | 41, 42, 48, 52, 26 |
pY572 | YLFFYRKSV | High | hTERT/mTERT | 6/6 | 48, 51, 64, 71, 38, 21 |
p865 | RLVDDFLLV | High | hTERT | 3/6 | 54, 49, 33 |
p988 | DLQVNSLQTV | Low | hTERT/mTERT | 6/6 | 46, 39, 48, 15, 19, 34 |
pY988 | YLQVNSLQTV | High | hTERT/mTERT | 6/6 | 61, 44, 79, 27, 33, 52 |
Peptides . | Sequence . | Relative avidity . | Protein of origin . | CTL response . | |
---|---|---|---|---|---|
R/T . | Percentage specific lysis . | ||||
p152 | LLARCALPV | Low | hTERT | 0/6 | — |
pY152 | YLARCALPV | High | hTERT | 0/6 | — |
p540 | ILAKFLHWL | High | hTERT | 5/6 | 64, 48, 57, 65, 41 |
p555 | ELLRSFFYV | Low | hTERT | 0/6 | — |
pY555 | YLLRSFFYV | High | hTERT | 1/6 | 16 |
p572 | RLFFYRKSV | Low | hTERT/mTERT | 5/6 | 41, 42, 48, 52, 26 |
pY572 | YLFFYRKSV | High | hTERT/mTERT | 6/6 | 48, 51, 64, 71, 38, 21 |
p865 | RLVDDFLLV | High | hTERT | 3/6 | 54, 49, 33 |
p988 | DLQVNSLQTV | Low | hTERT/mTERT | 6/6 | 46, 39, 48, 15, 19, 34 |
pY988 | YLQVNSLQTV | High | hTERT/mTERT | 6/6 | 61, 44, 79, 27, 33, 52 |
The HLA-A*0201-restricted hTERT-derived peptides and their respective relative avidity have been already described.29-31 HHD mice were immunized subcutaneously with 2 × 107 TU of recombinant lv-hTERT vector, and their splenocytes were restimulated in vitro 10 days later with each peptide: p540 or p865, or with the pool of native and optimized form of indicated cryptic peptides (p152/pY152, p555/pY55, p572/pY572, and p988/pY988; 10 μg/mL). Peptide-specific CTLs were tested against RMAS/HHD cells pulsed or not with the corresponding peptide. Data represent the percentage of specific lysis of peptide-pulsed target cells at a 50:1 splenocyte/target ratio, after subtraction of nonspecific lysis of nonpulsed target cells (0%-8%).
CTL indicates cytotoxic T lymphocytes; HHD, HLA-A2.1 transgenic mice; R/T, number of responder/total number of immunized; and —, not applicable.