Figure 1: TIMP-1 is expressed in an inducible manner on the surface of activated human and murine PMNs:
In A, human PMNs were activated 30 min at 37°C for with 10−8 M fMLP and then immunostained with non-immune rabbit IgG (top panel) or rabbit anti-TIMP-1 IgG (bottom panel) followed by goat anti-rabbit F(ab)2-conjugated to Alexa 488. Cells were examined using a Normarski objective (left panels) or confocal microscopy (right panels). The white bars are 5 microns in length. In B, human PMNs were incubated 30 min at 37°C with or without 10−8 M fMLP and then immunostained with rabbit anti-TIMP-1 IgG, rabbit anti-TIMP-2 IgG, or non-immune rabbit IgG followed by goat anti-rabbit F(ab)2-conjugated to Alexa 488. Images of immunostained cells were captured and analyzed using MetaMorph software. Data are mean + SEM (n = 150–200 cells/group). Data were analyzed using a One-Way ANOVA followed by pair-wise testing with two-tailed Student’s t-tests. Asterisk indicates P < 0.001 when compared to unstimulated cells stained for surface TIMP-1. In C, PMNs isolated from WT or Timp-1−/− mice were incubated at 37°C for 30 min with or without 10−6 M fMLP, and then immunostained with Alexa 488 for surface Timp-1. Data are mean + SEM; n = 150–200 cells/group. Data were analyzed using a One-Way ANOVA followed by pair-wise testing with two-tailed Student’s t-tests. Asterisk indicates P < 0.001 compared with unstimulated PMNs. The data shown are representative of at least 3 independent experiments.