Inflammatory cytokine responses of Met-5A PMC.

<p>A) Met-5A cell monolayers (n = 3 wells) were infected in triplicate wells with live D39 (MOI 1–200), stimulated with pure TNF-α (100ng/ml) or Nm-OM (100ng/ml) and also pre-stimulated with TNF-α (100ng/ml) for 4h and then infected with various MOI (1–200) of live D39. Control cells were left with medium alone. Cells were also treated with live D39 without any other stimulation (no cytokine production at any of the concentrations tested between MOI 1–200). After 24h, supernatants were removed and production of IL-6 and IL-8 quantified by ELISA. The bars represent mean cytokine levels and the errors bars the standard deviation from triplicate wells from a representative experiment (n = 2). * denotes statistically significant reduction in IL-6 or IL-8 compared to cells treated with TNFα only (p<0.05). B) Met-5A cells were challenged in triplicate wells with live D39 bacteria (1–100 MOI), TNFα (30 ng/monolayer) or Nm-OM (30ng/monolayer). At intervals up to 2h, cells were processed and NFkB p65 protein measured by ELISA. The columns and symbols denote the mean absorbance readings from n = 3 experiments and the error bars the standard errors of the mean. * denotes statistically significant increase compared to control cells (p<0.05). C) Met-5A cells were challenged in triplicate wells with live D39 bacteria (1–100 MOI), TNFα (30 ng/monolayer) or Nm-OM (30ng/monolayer). At intervals up to 2h, cells were processed and p38 MAP Kinase protein measured by ELISA. The columns and symbols denote the mean absorbance readings from n = 3 experiments and the error bars the standard errors of the mean. * denotes statistically significant increase compared to control cells (p<0.05).