FIGURE 2.

Identification and characterization of IL-2–responsive regulatory elements in the human granulysin 5′ flank. A, Transcriptional activity analysis in human primary CD4⁺ T cells. Transient transfection analysis of the pGL3−19,802/+62 or pGL3−329/+62 in primary CD4⁺ T cells was performed as described for B. B, Transient transfection analysis of granulysin 5′ (upper) and 3′ (lower) progressive flanking deletions. The genomic DNAs depicted on the left were used to control firefly luciferase expression in CRL-2105 T cells that were split 2 h after electroporation and cultured overnight for 16 h with and without IL-2. The data are given as the ratio of reporter activity of activated versus unactivated cells. DNA (2 μg) was used, comprising a 10:1 molar excess of the experimental plasmid over an internal control vector pRL–TK. The TK promoter-driven Renilla luciferase expression, which did not respond significantly to IL-2 in CRL-2105 T cells, served to correct for sample differences. In the absence of IL-2, the normalized expression of all granulysin promoter-bearing constructs was 10–40 times the levels obtained from the promoterless construct, with no reproducible correlation to any particular construct. C, Further characterization of the far-upstream enhancer of granulysin. Transient transfection analysis of the depicted DNAs in the context of the heterologous SV40 promoter was performed as described for B. Results are expressed as mean ± SEM. Data are representative of three independent experiments.