Figure 3

IκBα overexpression significantly inhibits rheumatoid T-cell-induced macrophage chemokine secretion of CXC, but not CC, chemokines. Using anti-CD3 labelled Dynabeads, synovial T cells were enriched from the mixed cell population obtained following enzymatic dissociation of synovial tissue samples from rheumatoid arthritis (RA) patients. Fixed RA T cells were cultured with macrophage-colony stimulating factor-differentiated monocytes infected with Ad0 and AdvIκBα at a T cell:monocyte ratio of 7:1 as described in Figure 2. 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 by ELISA. (a) Levels of chemokines for monocytes infected with Ad0 and AdIκBα (multiplicity of infection (MOI) 80:1) following stimulation with RA T cells. (b) GROα levels in uninfected, Ad0-infected (MOI 200:1) and AdvIκBα-infected (MOI 20:1, 40, 80:1 and 200:1) monocyte cultures when stimulated with RA T 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 AdIκBα-infected (as compared with Ad0-infected) cultures is indicated.