Figure 3.

(A–D) Feraspin R exhibits only limited toxicity to 868 TCR-transduced CTLs at the concentrations tested. 868 TCR-transduced CTLs were incubated for 24 hours with increasing doses of Feraspin R. (A) Live CD8⁺ T cells were gated, and apoptotic (annexin V⁺) and dead (allophycocyanin cytochrome 7⁺) cell gates were determined using a camptothecin-treated control for all doses of Feraspin R-treated CTLs. (B) Dead and apoptotic cells were identified and the population of annexin V-negative live cells were calculated as a percentage of total gated CD8⁺ cells (% living cells) in quadruplicate. (C) CTLs incubated for 24 hours with and without 1 mg/mL Feraspin R were washed and cultured for a further week. The percentage of living cells at days 1, 4, and 7 were determined in quadruplicate. (D) CTLs were incubated with Feraspin R in the presence of transfection reagents, and the percentage of living cells post-24-hour incubation was determined in duplicate.

Abbreviations: TCR, T-cell receptor; CTL, cytotoxic T lymphocyte; FITC, fluorescein isothiocyanate; CD, cluster of differentiation.