Table 1.

The Human Perforin 5′ Flank (−15,600 to +59) Promotes Regulated Reporter Expression in Transgenic Mice

Time	Medium	P	P + I	TCR	IL-2
h
0	1 (standard)	–	–	–	–
24	1	3	19	21	6
48	1	1	14	16	18

Splenic T cells of transgenic offspring (eight copies) were purified by magnetic microparticles conjugated to Thy1.2 mAb (MiniMACS™ system; Miltenyi Biotec). Luciferase was extracted from 10⁶ purified cells (>95% CD3⁺ and ∼40% CD8⁺) immediately and after culture for 24 and 48 h. Activations involved 10 ng/ml PMA (P) plus 1 μM ionomycin (P + I), cross-linking of CD3 (TCR; plate-bound 2C11 mAb), and 1,000 U/ml IL-2 (IL-2). Aliquots were measured in duplicate, and their average was used to calculate the fold inductions in reference to the activity obtained from cells prior to their in vitro culture (set as 1).