Journal:

Article Title: Endocarditis-Associated Oral Streptococci Promote Rapid Differentiation of Monocytes into Mature Dendritic Cells

doi: 10.1128/IAI.73.8.5015-5021.2005

Figure Lengend Snippet: Change in CD14 expression by monocytes after exposure to S. mutans. (A) PBMC control; (B) PBMC treated with S. mutans ATCC 25175 for 20 h. Monocytes were stained with CD14 and CD11c monoclonal antibodies. Note the reduction in CD14 when incubated with S. mutans.

Article Snippet: The MFI of CD86 clearly increased when PBMC were stimulated with M-CSF, IL-4 plus GM-CSF, L. casei , S , oralis ATCC 35037, S. mitis , S. mutans ATCC 2517), S. oralis ATCC 10557, S. sanguis , and S. mutans v1311 ( P < 0.05). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 3. caption a7 Effect of various bacteria on the MFI of CD86 expression by PBMC inside the monocyte gate.

Techniques: Expressing, Staining, Incubation

Journal:

Article Title: Endocarditis-Associated Oral Streptococci Promote Rapid Differentiation of Monocytes into Mature Dendritic Cells

doi: 10.1128/IAI.73.8.5015-5021.2005

Figure Lengend Snippet: Up-regulation of MHC class II expression by S. mutans-stimulated monocytes. PBMC from 3 individuals were cultured with two concentrations (105 and 106/ml) of S. mutans ATCC 25175 or A. viscosus for 20 h. Cells were then washed and stained with anti-human MHC class II monoclonal antibody. The vertical bars represent standard errors, and asterisks represent significant increases of the MFI of MHC class II labeling compared with the PBMC control. #, significant difference in MFI between S. mutans- and A. viscosus-stimulated cultures.

Article Snippet: The MFI of CD86 clearly increased when PBMC were stimulated with M-CSF, IL-4 plus GM-CSF, L. casei , S , oralis ATCC 35037, S. mitis , S. mutans ATCC 2517), S. oralis ATCC 10557, S. sanguis , and S. mutans v1311 ( P < 0.05). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 3. caption a7 Effect of various bacteria on the MFI of CD86 expression by PBMC inside the monocyte gate.

Techniques: Expressing, Cell Culture, Staining, Labeling

Journal:

Article Title: Endocarditis-Associated Oral Streptococci Promote Rapid Differentiation of Monocytes into Mature Dendritic Cells

doi: 10.1128/IAI.73.8.5015-5021.2005

Figure Lengend Snippet: CD83+ cells induced by S. mutans had a short life span. PBMC (0.5 × 106/0.5 ml) were stimulated with S. mutans ATCC 25175, A. viscosus, or P. intermedia (0.5 × 106/0.5 ml of enriched RPMI media that contains 1% penicillin and streptomycin). Cells were harvested in triplicate on days 1, 2, and 3 and labeled with CD83 and PI. The number of live CD83+ cells was calculated using the percentage of PI− CD83+ cells and the total cell count obtained using a hemacytometer. The vertical bars represent standard errors. The S. mutans data are representative of results from five different individuals, although in two cases, the peak number of viable DCs was found on day 2.

Article Snippet: The MFI of CD86 clearly increased when PBMC were stimulated with M-CSF, IL-4 plus GM-CSF, L. casei , S , oralis ATCC 35037, S. mitis , S. mutans ATCC 2517), S. oralis ATCC 10557, S. sanguis , and S. mutans v1311 ( P < 0.05). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 3. caption a7 Effect of various bacteria on the MFI of CD86 expression by PBMC inside the monocyte gate.

Techniques: Labeling, Cell Counting

Journal:

Article Title: Endocarditis-Associated Oral Streptococci Promote Rapid Differentiation of Monocytes into Mature Dendritic Cells

doi: 10.1128/IAI.73.8.5015-5021.2005

Figure Lengend Snippet: IL-12 p70 production by PBMC was CD14 dependent. PBMC (106/ml) were stimulated with S. mutans ATCC 25175 (106/ml) in the presence of neutralizing antibodies for TLR2, TLR4, or CD14 for 20 h in triplicate. IgG2a and IgG1 isotype controls were included for TLRs and CD14 antibodies, respectively. The vertical bars represent standard deviations. Asterisks represent significant decreases in IL-12 induction. This experiment is representative of the results from three experiments with three different individuals. Controls indicated that the anti-TLR2 and anti-TLR4 antibodies were effective under the conditions of these experiments. While anti-TLR2 failed to inhibit IL-12 production, it did inhibit the S. mutans (106/ml)-induced IL-10 response (from 731.3 ± 101.9 pg/ml to 272.4 ± 71.11 pg/ml; P < 0.01). Furthermore, anti-TLR4 inhibited IL-12 production induced in PBMC by LPS (E. coli, 100 ng/ml) from 491.8 ± 54.03 pg/ml to 77.26 ± 5.519 pg/ml (P < 0.01).

Article Snippet: The MFI of CD86 clearly increased when PBMC were stimulated with M-CSF, IL-4 plus GM-CSF, L. casei , S , oralis ATCC 35037, S. mitis , S. mutans ATCC 2517), S. oralis ATCC 10557, S. sanguis , and S. mutans v1311 ( P < 0.05). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 3. caption a7 Effect of various bacteria on the MFI of CD86 expression by PBMC inside the monocyte gate.

Techniques:

Journal:

Article Title: Endocarditis-Associated Oral Streptococci Promote Rapid Differentiation of Monocytes into Mature Dendritic Cells

doi: 10.1128/IAI.73.8.5015-5021.2005

Figure Lengend Snippet: Enhanced allogeneic mixed-lymphocyte reaction by S. mutans-stimulated monocytes. APC preparations were derived from overnight stimulation of PBMC with S. mutans ATCC 25175 (105 and 106/ml) or A. viscosus (105 and 106/ml). T-cell preparations from different individual PBMC were obtained using microbeads. Three concentrations of APC (103, 104, and 105/well) were cocultured with T cells (105/well) for 5 days. The cultures were then pulsed with [3H]thymidine (1 μCi/well) for 18 h, harvested, and counted for radioactivity (cpm) using Top Count. The T-cell control had 1,422 ± 1,284 cpm. The vertical bars represent standard errors. This experiment is representative of the results from two experiments with four different individuals.

Article Snippet: The MFI of CD86 clearly increased when PBMC were stimulated with M-CSF, IL-4 plus GM-CSF, L. casei , S , oralis ATCC 35037, S. mitis , S. mutans ATCC 2517), S. oralis ATCC 10557, S. sanguis , and S. mutans v1311 ( P < 0.05). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 3. caption a7 Effect of various bacteria on the MFI of CD86 expression by PBMC inside the monocyte gate.

Techniques: Derivative Assay, Radioactivity