Figure 2

Differential utilization of NFκB in activated-T-cell contact-dependent chemokine production by human macrophages. Macrophage-colony stimulating factor-differentiated monocytes were infected with AdIκBα or Ad0, an empty control virus. After a further 2 days of culture and replating, anti-CD3-activated T cells (Ttcr cells) and cytokine-activated T cells (Tck cells) were added at a lymphocyte:monocyte ratio of 7:1. After 18 hours, culture supernatants were isolated and levels of CC chemokines (monocyte chemoattractant protein 1 (MCP-1), macrophage inflammatory protein 1 alpha (MIP-1α), macrophage inflammatory protein 1 beta (MIP-1β), RANTES) and CXC chemokines (IL-8, growth-related gene product alpha (GROα) and interferon-gamma-inducible protein (IP-10)) were measured simultaneously by ELISA. (a) MIP-1α levels in uninfected, Ad0-infected (multiplicity of infection (MOI) 200:1) and AdIκBα-infected (MOI 40:1, 80:1 and 200:1) monocyte cultures when stimulated with Ttcr-cells or Tck-cells. (b) and (c) Levels of CC and CXC chemokines in Ad0-infected and AdIκBα-infected monocytes (MOI 80:1) following stimulation with (b) Ttcr cells and (c) Tck cells. Data represent the mean of triplicate cultures ± standard deviation and are representative of at least three experiments. Statistically significant reduction in chemokine levels in AdvIκBα-infected (as compared with Ad0-infected) cultures is indicated.