Media conditioned by IM MNCs have a greater ability to cleave VCAM-1 and c-kit. (A) MNCs were incubated in serum-free IMDM at 37°C and 5% CO2 for 24 hours. Recombinant rhuVCAM-1 was incubated for 15 minutes at 37°C in the presence of medium conditioned by PB mononuclear cells from either healthy volunteers (n = 8) or patients with IM (n = 8) or in the presence of purified human NE, MMP-9, or PBS. Following preincubation, samples were analyzed by Western blot with a goat anti–human VCAM-1 antibody. The arrows represent the fragments of the extracellular domain of VCAM-1. (B) Purified extracellular domains of rhuc-kit was incubated at 37°C for 6 hours in the presence of medium conditioned by PB mononuclear cells from either healthy volunteers (n = 8) or patients with IM (n = 8) or in the presence of purified human NE, MMP-9, or PBS. Following preincubation, samples were analyzed by Western blot with a goat anti–human c-kit antibody. The arrows represent the fragments of the extracellular domain of c-kit.
