Antigen-specific reactivity in peripheral blood mononuclear cells from patients immunized with peptide-pulsed dendritic cells
| Patient no. . | Antigens . | % of IFN-γ-producing lymphocytes* . | In vitro CTL response (% specific lysis)† . | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| HIV . | E75 . | GP2 . | HIV . | M1.1 . | M1.2 . | HIV . | E75 . | GP2 . | HIV . | M1.1 . | M1.2 . | ||
| 1SR | HER-2/neu | 0.1 | 0.3 | 0.1 | 11 | 19 | 16 | ||||||
| 2ZP | HER-2/neu | 0.1 | 0.9 | 0.1 | 19/15 | 49/17 | 26/19‡ | ||||||
| 3SJ | HER-2/neu | 0.1 | 0.2 | 0.3 | 9 | 19 | 8 | ||||||
| 4KG | HER-2/neu | 0.2 | 0.3 | 0.1 | nd | ||||||||
| 5TG | HER-2/neu | 0.2 | 0.2 | 0.3 | 6 | 4 | 0 | ||||||
| 6HA | HER-2/neu | 0.1 | 1.2 | 0.3 | 9 | 37/11 | 6 | ||||||
| 7WL | MUC1 | 0.1 | 0.7 | 0.9 | 15/19 | 33/9 | 59/24 | ||||||
| 8MI | MUC1 | 0.1 | 0.8 | 2 | 12/14 | 49/21 | 68/17 | ||||||
| 9KI | MUC1 | 0.3 | 0.1 | 0.2 | 18 | 17 | 13 | ||||||
| 10HA | MUC1 | 0.1 | 0.5 | 1.6 | 11/8 | 28/8 | 58/19 | ||||||
| Patient no. . | Antigens . | % of IFN-γ-producing lymphocytes* . | In vitro CTL response (% specific lysis)† . | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| HIV . | E75 . | GP2 . | HIV . | M1.1 . | M1.2 . | HIV . | E75 . | GP2 . | HIV . | M1.1 . | M1.2 . | ||
| 1SR | HER-2/neu | 0.1 | 0.3 | 0.1 | 11 | 19 | 16 | ||||||
| 2ZP | HER-2/neu | 0.1 | 0.9 | 0.1 | 19/15 | 49/17 | 26/19‡ | ||||||
| 3SJ | HER-2/neu | 0.1 | 0.2 | 0.3 | 9 | 19 | 8 | ||||||
| 4KG | HER-2/neu | 0.2 | 0.3 | 0.1 | nd | ||||||||
| 5TG | HER-2/neu | 0.2 | 0.2 | 0.3 | 6 | 4 | 0 | ||||||
| 6HA | HER-2/neu | 0.1 | 1.2 | 0.3 | 9 | 37/11 | 6 | ||||||
| 7WL | MUC1 | 0.1 | 0.7 | 0.9 | 15/19 | 33/9 | 59/24 | ||||||
| 8MI | MUC1 | 0.1 | 0.8 | 2 | 12/14 | 49/21 | 68/17 | ||||||
| 9KI | MUC1 | 0.3 | 0.1 | 0.2 | 18 | 17 | 13 | ||||||
| 10HA | MUC1 | 0.1 | 0.5 | 1.6 | 11/8 | 28/8 | 58/19 | ||||||
PBMNCs indicate peripheral blood mononuclear cells; CTL, cytotoxic T lymphocytes; nd, not done.
PBMNCs from each patient were incubated with autologous PBMNCs pulsed with the indicated peptides for 6 hours and IFN-γ production was assessed by flow cytometry after intracellular staining for IFN-γ in CD8 positive lymphocytes. Numbers represent the percentage of IFN-γ-expressing cells in the lymphocyte gate.
PBMNCs from each patient were incubated with peptide-pulsed autologous DCs as indicated. After 7 days the cells were restimulated with irradiated peptide-pulsed PBMNCs. The cytotoxic activity of the CTL was determined 5 days later in a standard 57Cr-release assay. The data are presented at E:T of 30:1.
Lysis of targets cells pulsed with the cognate/irrelevant peptide.