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Rapid Tissue Factor Induction by Oral Streptococci and Monocyte-IL-1ß

¹ Department of Endodontics, School of Dentistry, Lyons Building, Rm. 441, 520 N. 12th Street, PO Box 980566, Richmond, VA 23298-0566, USA;
² Department of Biostatistics and
³ Department of Microbiology and Immunology, School of Medicine, Virginia Commonwealth University, Richmond, VA, USA

View larger version (8K): [in this window] [in a new window]   Figure 1. Tissue factor induction by oral streptococci in a transwell system. PBMC (106) were stimulated with 4 strains of oral streptococci (107) in 200 µL enriched RPMI without antibiotics in transwell inserts. The up-regulation of endothelial TF activity in 6 hrs was measured with the factor Xa assay (mU/mL). Data from 4 independent experiments were analyzed with ANOVA. *Significant increase of TF activity when compared with PBMC control by Tukey’s HSD (p < 0.005). The vertical bars represent standard errors.

View larger version (8K): [in this window] [in a new window]   Figure 2. IL-1ß neutralizing antibodies block the induction of TF activity. PBMC (106/mL) were challenged with live S. mutans (107/mL) for 4 hrs. Supernatant fluids (SF control) were then harvested and filtered with 0.2 µ to remove bacteria. Endothelial TF activity induced by SF control in the presence of neutralizing antibodies (anti-IL-1, anti-IL-1ß, anti-IL1-ra, anti-TNF-) was measured with the factor Xa assay (mU/mL). Supernatant fluids from LPS-challenged (100 ng/mL) PBMC cultures were included as a positive control. Data from 4 independent experiments were analyzed with a mixed ANOVA. Controls with rIL-1ß (10 ng/mL) induced comparable factor Xa activity (5–15 mU/mL). *Significantly lower factor Xa titer than SF control by Tukey’s HSD (p < 0.05). The vertical bars represent standard errors.

View larger version (12K): [in this window] [in a new window]   Figure 3. Monocytes are the major cellular source for endothelial TF activity. PBMC and monocyte-depleted PBMC (Mo-depleted) preparations (106/mL) in triplicate were challenged with live S. mutans, S. oralis, and S. sanguinis 10556 at 107/mL for 4 hrs. The supernatant fluids were harvested and assayed for their induction of endothelial TF activity (factor Xa). Data were analyzed with two-way ANOVA, and the differences between PBMC and Mo-depleted preparations for each strain were examined with post hoc comparisons. This experiment was repeated with another donor with a similar pattern. *Significantly lower factor Xa titer than its respective PBMC control (p < 0.0002). The vertical bars represent standard deviations.