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. Author manuscript; available in PMC: 2014 Sep 15.

Published in final edited form as: Clin Cancer Res. 2013 Jul 9;19(18):5079–5091. doi: 10.1158/1078-0432.CCR-13-0955

Antigen-specific CTLs of patient #1032 showed recognition of TAA after the initial stimulation (A), as well as recognition of PRAME and WT1 after the 3^rd stimulation (B) in IFNγ-ELISpot (mean ± SD). (C) Cytolytic activity in a standard ⁵¹Cr-release assay against antigen-pulsed autologous target cells (PHA-blasts, PHAB) at E:T ratios from 40:1 to 1.2:1. (D) Percent of remaining leukemia cells in co-culture of CTLs with autologous leukemia blasts on day 1 and 3 in presence of IL2 (50U/ml) at an E:T ratio of 10:1. Leukemic cells were detected by co-staining with CD10 and CD19 antibodies, T cells by staining with CD3 antibody; nonspecific T cells (white bars) were used as control for unspecific lysis (CTLs: black bars). (E) Assessment of absolute leukemia cell counts in co-culture with CTLs (black line) and nonspecific T cells (dashed line) at an E:T ratio of 10:1 by FACS analysis using counting beads. (F) Co-culture at a 1:1 ratio of autologous leukemia cells to CTLs (black bar) or nonspecific T cells (white bar) in IFNγ-ELISpot assay (mean ± SD). (G) Immunohistochemistry of leukemia blasts.

Antigen-specific CTLs of patient #1032 showed recognition of TAA after the initial stimulation (A), as well as recognition of PRAME and WT1 after the 3^rd stimulation (B) in IFNγ-ELISpot (mean ± SD). (C) Cytolytic activity in a standard ⁵¹Cr-release assay against antigen-pulsed autologous target cells (PHA-blasts, PHAB) at E:T ratios from 40:1 to 1.2:1. (D) Percent of remaining leukemia cells in co-culture of CTLs with autologous leukemia blasts on day 1 and 3 in presence of IL2 (50U/ml) at an E:T ratio of 10:1. Leukemic cells were detected by co-staining with CD10 and CD19 antibodies, T cells by staining with CD3 antibody; nonspecific T cells (white bars) were used as control for unspecific lysis (CTLs: black bars). (E) Assessment of absolute leukemia cell counts in co-culture with CTLs (black line) and nonspecific T cells (dashed line) at an E:T ratio of 10:1 by FACS analysis using counting beads. (F) Co-culture at a 1:1 ratio of autologous leukemia cells to CTLs (black bar) or nonspecific T cells (white bar) in IFNγ-ELISpot assay (mean ± SD). (G) Immunohistochemistry of leukemia blasts.