Fig. 5.

Responsiveness of CTLs correlates with lck expression, and treatment with Z-LL-H prevents the development of AINR in specific CTLs. (A) BK bulk CTLs were stimulated during the indicated periods of time with C1R/A11 cells preincubated with the indicated synthetic peptides. The expression of lck was accessed by immunoblotting and quantified as described above. (B) CTLs preactivated by using the indicated peptides were restimulated with IVT-pulsed APCs after 48 h, and their proliferation was evaluated by a [³H]thymidine incorporation assay. Shown are the results of one representative of two comparable experiments. (C and D) BK bulk CTLs were activated for 2 h on plastic plates with absorbed CD3-specific Ab, transferred to a clean plate, and cultured for 48 h either in the absence or presence of 20 μM Z-LL-H. Lck expression in these cells was evaluated by immunoblotting. Shown are data of one representative experiment. (D) Cells were then activated by IVT-peptide-pulsed APCs, and their proliferation was evaluated by a [³H]thymidine incorporation assay. The results are expressed as the percentage relative to thymidine incorporation in control cultures not stimulated by anti-CD3 and cultured either with or without Z-LL-H. Shown are the means ± SD of three experiments.