Specific recognition of HLA-A0201 binding fibromodulin-derived peptides present on CD40L-stimulated CLL cells by autologous T cells
. | . | FMOD-specific T cells (per 100 000) by ELISPOT . | . | . | . | . | . | ||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| . | . | Native autologous T cells . | T-cell expansion without pulsing the APCs . | APCs were pulsed with F1/2 or F3/4 to expand T cells . | . | . | FMOD-specific CD8+ T cells by dimer-peptide staining (in %) . | ||||
| Patient . | FMOD peptides . | . | . | Elispot: APCs pulsed with F1/2 or F3/4 . | Elispot: APCs pulsed with F1 or F3 . | Elispot: APCs pulsed with F2 or F4 . | . | ||||
| CLL-1 | F3/4 | 12 | 111 | 140 | 215 | 420 | 15.6 | ||||
| CLL-2 | F1/2 | 4 | 54 | 160 | 22 | 8 | 30.4 | ||||
| F3/4 | 4 | 20 | 42 | 26 | 56 | ND | |||||
| CLL-3 | F3/4 | 0 | ND | 108 | ND | ND | ND | ||||
| CLL-4 | F1/2 | 230 | 884 | 1303 | 1228 | 1164 | 12.2 | ||||
| F3/4 | 414 | 388 | 976 | 0 | 560 | ND | |||||
| CLL-7 | F1/2 | 0 | ND | 24 | ND | ND | ND | ||||
| F3/4 | 0 | ND | 4 | ND | ND | ND | |||||
| CLL-8 | F1/2 | 2 | 60 | 740 | 470 | 360 | ND | ||||
| F3/4 | 0 | 40 | 68 | 0 | 88 | 6.1 | |||||
. | . | FMOD-specific T cells (per 100 000) by ELISPOT . | . | . | . | . | . | ||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| . | . | Native autologous T cells . | T-cell expansion without pulsing the APCs . | APCs were pulsed with F1/2 or F3/4 to expand T cells . | . | . | FMOD-specific CD8+ T cells by dimer-peptide staining (in %) . | ||||
| Patient . | FMOD peptides . | . | . | Elispot: APCs pulsed with F1/2 or F3/4 . | Elispot: APCs pulsed with F1 or F3 . | Elispot: APCs pulsed with F2 or F4 . | . | ||||
| CLL-1 | F3/4 | 12 | 111 | 140 | 215 | 420 | 15.6 | ||||
| CLL-2 | F1/2 | 4 | 54 | 160 | 22 | 8 | 30.4 | ||||
| F3/4 | 4 | 20 | 42 | 26 | 56 | ND | |||||
| CLL-3 | F3/4 | 0 | ND | 108 | ND | ND | ND | ||||
| CLL-4 | F1/2 | 230 | 884 | 1303 | 1228 | 1164 | 12.2 | ||||
| F3/4 | 414 | 388 | 976 | 0 | 560 | ND | |||||
| CLL-7 | F1/2 | 0 | ND | 24 | ND | ND | ND | ||||
| F3/4 | 0 | ND | 4 | ND | ND | ND | |||||
| CLL-8 | F1/2 | 2 | 60 | 740 | 470 | 360 | ND | ||||
| F3/4 | 0 | 40 | 68 | 0 | 88 | 6.1 | |||||
T cells from CLL patients were expanded using autologous, CD40L-stimulated CLL cells as APCs. IFN-γ-ELISPOT assays were performed on days 14 or 21 with autologous CD40L-stimulated CLL cells as APCs. HLA-A0201 binding fibromodulin-derived peptides were added to obtain the number of fibromodulin-specific spots. Either a mix of F1/2 or F3/4 or the single peptides were added. Coincubation of CD40L-stimulated CLL cells with DMSO and autologous T cells served as negative controls, and the number of spots detected were subtracted from the experimental values. Given are the numbers of fibromodulin-specific spots per 100 000 T cells. The precursor T-cell frequency was investigated by coincubation of CD40L-stimulated CLL cells and unstimulated autologous T cells together with the fibromodulin-derived peptides (F1/2 or F3/4). The percentage of CD8+ T cells on day 28 recognizing fibromodulin-derived peptides bound to HLA-A2—dimer is given after subtraction of the negative control HLA-A2—dimer coincubated with DMSO. T cells were obtained using CD40L-stimulated fibromodulin-pulsed CLL cells as APCs.