Figure 2.

HZ and trophozoite phagocytosis, and rhIL-1beta enhance MMP-9 protein expression (in cell lysates) and enzyme activity (in cell supernatants) in human adherent monocytes. Abrogation of the HZ effect by anti-hIL-1beta antibodies. Human adherent monocytes were unfed, fed with HZ or latex particles treated or not with rhIL-1beta (20 ng/ml) or blocking anti-hIL-1beta antibodies (30 ng/ml) as indicated. Panel A. Western blot with anti-MMP-9 antibodies and densitometric quantification of MMP-9 protein. After 3 h phagocytosis and a further incubation during 48 h, cell lysates were prepared, separated by PAGE (8% polyacrylamide) blotted and probed with anti-MMP-9 monoclonal antibodies (1/1000 final dilution). The 92-kDa band in the gel corresponds to pro-MMP-9. Data are given as arbitrary densitometric units (mean values ± SD of four independent experiments). Panel B. Gelatin zymography and densitometric quantification of MMP-9 enzyme activity. After 3 h phagocytosis and a further incubation during 48 h, cell supernatants were separated by PAGE (8% polyacrylamide gel containing 0.1% gelatin) under non-denaturing and non-reducing conditions. The 83-kDa negative bands in the gel correspond to MMP-9 enzyme activity. Data are given as arbitrary densitometric units (mean values ± SD of four independent experiments). Data (Panel A, Panel B) were analysed for significance by Student's t-test. Significance of differences (column/lane numbers): HZ-fed(7)/rhIL1beta(2)-stimulated vs control(1)/anti-hIL1beta-stimulated(3)/latex-fed(4) monocytes, p < 0.01 (Panel A) or p < 0.05 (Panel B). rhIL1beta-stimulated(2,5,8) vs HZ-fed(7,8) monocytes, n.s.; anti-hIL1beta-stimulated(3,6,9)/latex-fed(4) vs unfed(1) monocytes, n.s.