Figure 7.

Transmigrated monocytes express enhanced costimulatory molecules. Purified populations of monocytes (10⁷) were cultured with IFN-γ–treated HUVEC monolayers in the upper chamber of a 8-μm-pore polycarbonate transwell. (A) Monocyte CD86 protein expression was determined by FACS^® 24 h after transmigration. Illustrated is the FACS^® analysis of CD86 in resting monocytes and transmigrated monocytes. Isotype control PE antibody served as control. Representative analysis of three experiments. (B) Monocytes were harvested from the lower chamber after 12 h. Illustrated is the expression of IL-1α by RNase protection from resting, transmigrated monocytes and positive control IFN-γ–activated monocytes. (C) RNA was harvested from cocultures of monocytes and IFN-γ–activated endothelial cells. Illustrated is the expression of IL-12 by RNase protection in resting monocytes, monocytes cocultured with IFN-γ–activated ECs, and LPS (1 μg/ml)-stimulated monocytes. Unactivated or IFN-γ–activated ECs do not express IL-12, and IFN-γ alone only weakly induces IL-12 expression in resting monocytes (not shown).