TIMP-1 and IL-10 inhibit apoptosis in JD38 cells.
(A) JD38 cells (3 × 105 cells/mL) were incubated for 48 hours in serum-free media with 0 to 25 ng/mL recombinant IL-10, and viability was determined. (B) JD38 cells (3 × 105cells/mL) were incubated for 48 hours in serum-free media with 0 to 250 ng/mL recombinant IL-10. Apoptosis (░) and proliferation (▪) were determined. (C) JD38 cells and TIMP-1 clone 24 (3 × 105cells/mL) were incubated for 48 hours in serum-free media and viability was determined. Viability is shown on the y-axis. JD38: JD38 cells incubated in serum-free media. JD38 + TIMP-1: JD38 cells incubated with 143 nM rTIMP-1. JD38 + IL-10: JD38 cells incubated with 24 ng/mL rIL-10. Clone 24: TIMP-1–expressing JD38 clone. 24 + aTIMP-1: TIMP-1–expressing clone 24 was incubated with 1 μg/mL neutralizing anti–TIMP-1 monoclonal antibody. 24 + aIL-10: IL-10 was inactivated in TIMP-1–expressing clone 24 by incubation with neutralizing antibodies to IL-10 (0.5 μg/mL) and with 50 pg/mL soluble IL-10 receptor. Incubation with equivalent quantity of isotype control antibody protein was performed in each experiment.
