Figure 1.
Figure 1. Frequencies and functional activity of MUC1-specific CD8 T cells in PB and BM of MM patients or healthy donors. (A) One representative dot plot from a patient's PB showing tetramer binding cells among PI- mononuclear cells (2.5% tetramer binding cells among CD8+ T cells). (B) Frequencies of MUC1-specific CD8 T cells indicated as percent of total CD8 T cells from each individual patient (P; n = 50) and healthy donor (HD; n = 14) analyzed. The solid line indicates the maximum frequency measured in the group of healthy donors. The dotted line indicates maximum frequency of CD8 T cells binding HIV tetramers (n = 7). Significant differences between myeloma samples and corresponding samples from healthy donors are depicted for PB (†, P < .02) and BM (*, P < .05). Values from patients with elevated numbers of MUC1 specific CD8 T cells in either PB or BM are indicated by interrupted lines connecting corresponding values of PB and BM. As control, frequencies of the patient's CD8 T cells binding irrelevant tetramers containing HIV-derived peptide SLYNTVATL are shown (HIV). (C) Frequencies of IFN-γ secreting MUC1 peptide–reactive memory T cells in PB (▴; n = 10) and BM (⬡; n = 19) of altogether 21 myeloma patients (P) and of BM samples of 5 healthy donors (HD) (▵). Numbers of MUC1-reactive cells were calculated as difference between mean spot numbers in test wells and control wells and indicated as numbers of 106 purified T cells. †, overall significant difference between BM samples from myeloma patients and healthy donors (P < .05). *, overall significant difference between PB and BM of myeloma patients (P < .01). Values from patients with elevated numbers of MUC1-reactive T cells in either PB or BM are indicated by dotted lines connecting corresponding values of PB and BM. §, overall significant difference between PB and BM in paired samples from patients containing MUC1-reactive memory T cells (P < .02). (D) Percent of MUC1-specific lysis quantifies the significant difference between specific lysis of LLLLTVLTV-loaded T2 cells and lysis of T2 cells loaded with control peptide HLVEALYLV measured at an effector-target ratio of 25:1. No MUC1-specific cytotoxicity was observed in PB T cells at different effector-target ratios (6:1 vs 50:1). *, overall significant difference between cytotoxicity of BM T cells compared to PB T cells (P < .03). (E) Percent of perforin content of CD45RA- (⬡) and CD45RA+ CD8 T cells (○) from PB and BM samples of 3 myeloma patients after 7 days of coculture with autologous DCs pulsed with MUC1-derived peptide LLLLTVLTV. *, significant difference between perforin content of BM T cells compared to PB T cells (P = .04).

Frequencies and functional activity of MUC1-specific CD8 T cells in PB and BM of MM patients or healthy donors. (A) One representative dot plot from a patient's PB showing tetramer binding cells among PI- mononuclear cells (2.5% tetramer binding cells among CD8+ T cells). (B) Frequencies of MUC1-specific CD8 T cells indicated as percent of total CD8 T cells from each individual patient (P; n = 50) and healthy donor (HD; n = 14) analyzed. The solid line indicates the maximum frequency measured in the group of healthy donors. The dotted line indicates maximum frequency of CD8 T cells binding HIV tetramers (n = 7). Significant differences between myeloma samples and corresponding samples from healthy donors are depicted for PB (†, P < .02) and BM (*, P < .05). Values from patients with elevated numbers of MUC1 specific CD8 T cells in either PB or BM are indicated by interrupted lines connecting corresponding values of PB and BM. As control, frequencies of the patient's CD8 T cells binding irrelevant tetramers containing HIV-derived peptide SLYNTVATL are shown (HIV). (C) Frequencies of IFN-γ secreting MUC1 peptide–reactive memory T cells in PB (▴; n = 10) and BM (⬡; n = 19) of altogether 21 myeloma patients (P) and of BM samples of 5 healthy donors (HD) (▵). Numbers of MUC1-reactive cells were calculated as difference between mean spot numbers in test wells and control wells and indicated as numbers of 106 purified T cells. †, overall significant difference between BM samples from myeloma patients and healthy donors (P < .05). *, overall significant difference between PB and BM of myeloma patients (P < .01). Values from patients with elevated numbers of MUC1-reactive T cells in either PB or BM are indicated by dotted lines connecting corresponding values of PB and BM. §, overall significant difference between PB and BM in paired samples from patients containing MUC1-reactive memory T cells (P < .02). (D) Percent of MUC1-specific lysis quantifies the significant difference between specific lysis of LLLLTVLTV-loaded T2 cells and lysis of T2 cells loaded with control peptide HLVEALYLV measured at an effector-target ratio of 25:1. No MUC1-specific cytotoxicity was observed in PB T cells at different effector-target ratios (6:1 vs 50:1). *, overall significant difference between cytotoxicity of BM T cells compared to PB T cells (P < .03). (E) Percent of perforin content of CD45RA- (⬡) and CD45RA+ CD8 T cells (○) from PB and BM samples of 3 myeloma patients after 7 days of coculture with autologous DCs pulsed with MUC1-derived peptide LLLLTVLTV. *, significant difference between perforin content of BM T cells compared to PB T cells (P = .04).

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