Figure 5 - Neutralizing anti-S100A8 and anti-S100A9 antibodies block the migration of macrophages and tumour cells to lungs.

(a) Phosphorylation of p38 MAP kinase stimulated by S100A8–GST, GST, S100A8–GST–LCM or GST–LCM in macrophages (M) and LLC cells. Cells were treated for 5–30 min and analysed by western blotting. SB203580 (SB) or anti-S100A8 antibodies suppressed phosphorylation of p38 stimulated by S100A8–GST. The intensity of the bands in phospho-p38 was normalized to those in p38 and the relative values are shown under panels. (b) The inhibition of p38 signalling suppressed migration of LLC cells in response to S100A8. S100A8–GST (100 pg ml^-1), GST (100 pg ml^-1), SB203580 (2 M) in DMSO, or an equal volume of DMSO (D, control) was added to lower wells (n = 5). Means s.d. are shown. (c) The numbers of Mac 1⁺-myeloid cells in premetastatic lungs from LLC-bearing or B16-bearing mice, which were treated with control IgG, anti-S100A8 antibody alone (anti-A8), or both anti-S100A8 and anti-S100A9 (anti-A9) antibodies. Means s.d. are shown (n = 5, asterisk indicates P <0.05). (d) The numbers of rhodamine-labelled tumour cells in lungs from normal (N), LLC-bearing or B16-bearing mice, which were treated with control IgG, anti-S100A8 antibody alone, or both anti-S100A8 and anti-S100A9 antibodies before the injection of these cells. Detection was carried out at 5 h and 24 h after the injection of tumour cells into the tail vein. Means s.d. are shown (n = 10, asterisk indicates P <0.05).