Figure 3. The ectopically expressed survivin-specific TCR is functional and specific, but not fratricidal, in vitro.

(A–C, E, and F) Symbols represent the mean of triplicates per donor; error bars represent the mean ± SD. (A) Production of IFN-γ (ELISpot) in response to LML and ELT peptides by nontransduced (NT, black circles) and transduced (TD, white squares) T cells (n = 5). (B) Killing of LML-pulsed T2 cells (⁵¹Cr release) by NT and TD T cells, with the percentage of specific lysis at an E/T ratio of 20:1 (n = 3). (C) Evaluation of HLA restriction of the killing of LML-pulsed T2 cells by preincubation with HLA class I–blocking Ab (dotted line) or HLA class II–blocking Ab (dashed line), or in the absence of Ab (solid line) by TD T cells (white squares) and NT T cells (black squares, solid line). Data are representative of 2 independent experiments. (D) Expansion of transgenic T cells with weekly antigen-specific stimulations generated from HLA-A*02⁺ (black circles, solid line) or HLA-A*02^– (white squares, dashed line) donors. Mean ± SD, n = 7, P = NS. (E and F). ⁵¹Cr-release assays of NT (black circles) and TD T cells (white squares) against activated HLA-A*02⁺ target T cells loaded or not with LML or ELT peptide. Percentage of specific lysis at an E/T ratio of 20:1 of survivin-specific TCR expressed in (E) HLA-A*02^– (n = 7) or (F) HLA-A*02⁺ donors (n = 7).