TABLE 2

The effect of TMI-1 on the inhibition of TNF-α secretion in cell-based assays

The cell-based activity of TMI-1 was determined in murine and human monocyte cell lines, Raw cells, THP-1, human primary monocytes, and human whole blood for the inhibition of LPS or zymosan-induced TNF-α secretion. Cells were pretreated with TMI-1 or vehicle control at various concentrations for 1 h before addition of LPS or Zymosan. The cells were cultured for 4 h, and the supernatants were collected and frozen at −80°C. The concentration of soluble TNF-α was determined by an ELISA assay. In our systems, the range of soluble TNF-α induced by LPS (10 ng/ml) is 10 to 50 ng/ml in Raw cells, 5 to 15 ng/ml in human whole blood, and 2 to 10 ng/ml in human monocytes. In THP-1 cells, the range of soluble TNF-α induced by LPS (10 μg/ml) is 0.5 to 5 ng/ml. The range of soluble TNF-α induced by Zymosan (10 μg/ml) is 5 to 15 ng/ml in human whole blood. The data are averages from three experiments.