Journal: Molecular Medicine Reports

Article Title: Expression of programmed death 1 ligand 1 on periodontal tissue cells as a possible protective feedback mechanism against periodontal tissue destruction

doi: 10.3892/mmr.2016.4824

Figure Lengend Snippet: Inducible expression of PD-L1 on PDLCs by inflammatory cytokines. (A) Flow cytometry histogram overlays of PDLCs stimulated with a series of inflammatory cytokines. (B) Comparison of the induced expression of PD-L1. Data is presented as the mean ± standard error of the mean of three independent experiments. All experiments showed a significant increase in expression levels of PD-L1 ( * P<0.05, vs. control). The TNF-α and the combined cytokine groups induced significantly higher expression levels of PD-L1, compared with the other cytokines assessed ( † P<0.05).

Article Snippet: The sections were incubated with rabbit anti-mouse PD-L1 polyclonal antibodies (dilution, 1:200; Abcam, Cambridge, MA, USA; cat. no. ab58810) overnight at 4°C, followed by incubation with biotinylated goat anti-rabbit secondary antibodies (1:5,000; ZSGB-BIO, Beijing, China) for 30 min at room temperature, and with streptavidin-peroxidase complex solution for another 30 min at room temperature.

Techniques: Expressing, Flow Cytometry

Journal: Molecular Medicine Reports

Article Title: Expression of programmed death 1 ligand 1 on periodontal tissue cells as a possible protective feedback mechanism against periodontal tissue destruction

doi: 10.3892/mmr.2016.4824

Figure Lengend Snippet: Common periodontal pathogens induce the expression of PD-L1 on PDLCs. (A) Flow cytometry histogram overlays of PDLCs co-cultured with P.g , F.n and P.i . (B) Comparison of expression levels of PD-L1 induced by P.g, F.n and P.i. Data are expressed as the mean ± standard error of the mean of three independent experiments. All three strains significantly increased the expression of PD-L1 on the PDLCs, * P<0.05. P.g, Porphyromonas gingivalis ; F.n, Fusobacterium nucleatum ; P.i, Prevotella intermedia ; PD-L1, programmed death 1 ligand 1; PDLCs, periodontal ligament cells.

Article Snippet: The sections were incubated with rabbit anti-mouse PD-L1 polyclonal antibodies (dilution, 1:200; Abcam, Cambridge, MA, USA; cat. no. ab58810) overnight at 4°C, followed by incubation with biotinylated goat anti-rabbit secondary antibodies (1:5,000; ZSGB-BIO, Beijing, China) for 30 min at room temperature, and with streptavidin-peroxidase complex solution for another 30 min at room temperature.

Techniques: Expressing, Flow Cytometry, Cell Culture

Journal: Molecular Medicine Reports

Article Title: Expression of programmed death 1 ligand 1 on periodontal tissue cells as a possible protective feedback mechanism against periodontal tissue destruction

doi: 10.3892/mmr.2016.4824

Figure Lengend Snippet: TNF-α- and IFN-γ-induced expression of PD-L1 on PDLCs caused apoptosis of activated T cells. (A) Two-color flow cytometry histograms of activated PBMCs co-cultured with untreated PDLCs. PDLCs pretreated with (B) IFN-γ or (C) TNF-α, or incubated with (D) TNF-α-pretreated PDLCs and anti-PD-L1 antibody. (E) PI − cells were gated in R1 to exclude necrotic cells. Comparison of (F) CD8 + and (G) CD4 + T cell apoptosis induced by TNF-α and IFN-γ, and in the presence of anti-PDL1 antibodies. Data are presented as the mean ± standard error of the mean of three independent experiments. The increases in T cell apoptosis caused by pretreatment with TNF-α or IFN-γ were statistically significant ( * P<0.05), and the presence of anti-PD-L1 antibodies caused a considerable reduction in the fraction of apoptotic T cells ( † P<0.05). PD-L1, programmed death 1 ligand 1; PDLCs, periodontal ligament cells; IFN-γ, ibterferon-γ; TNF-α, tumor necrosis factor-α; APC, antigen-presenting cell; PI, propidium iodide; FITC, fluorescein isothiocyanate.

Article Snippet: The sections were incubated with rabbit anti-mouse PD-L1 polyclonal antibodies (dilution, 1:200; Abcam, Cambridge, MA, USA; cat. no. ab58810) overnight at 4°C, followed by incubation with biotinylated goat anti-rabbit secondary antibodies (1:5,000; ZSGB-BIO, Beijing, China) for 30 min at room temperature, and with streptavidin-peroxidase complex solution for another 30 min at room temperature.

Techniques: Expressing, Flow Cytometry, Cell Culture, Incubation

Journal: Molecular Medicine Reports

Article Title: Expression of programmed death 1 ligand 1 on periodontal tissue cells as a possible protective feedback mechanism against periodontal tissue destruction

doi: 10.3892/mmr.2016.4824

Figure Lengend Snippet: Expression of PD-L1 on PDLCs improves survival of PDLCs. Flow cytometry histrograms of (A) PDLCs, (B) PHA-activated PBMCs, (C) PDLCs co-cultured with activated PBMCs, (D) PDLCs pretreated with TNF-α and co-cultured with activated PBMCs, and (E) PDLCs pretreated with TNF-α, and incubated with activated PBMCs and anti-PD-L1 antibodies. (F) Comparison of PDLC survival, according to the percentages of CFSE + /PI − cells. a, c, d and e represent the PDLC control, untreated control, TNF-α induced and TNF-α induced+anti-PD-L1 groups, respectively. Data are expressed as the mean ± standard error of the mean of three independent experiments. Co-culturing the activated PMBCs with untreated PDLCs resulted in a significant decrease in viable PDLCs ( * P<0.05). † P<0.05, significant increase in the viability of the PDLCs following pretreatment with TNF-α. The addition of anti-PD-L1 antibody caused a significant decrease in the viability of the PDLCs pretreated with TNF-α ( ‡ P<0.05). PD-L1, programmed death 1 ligand 1; PDLCs, periodontal ligament cells; PMBCs, peripheral blood mononuclear cells; PI, propidium iodide; CFSE, carboxyfluorescein diacetate succinimidyl ester; TNF-α, tumor necrosis factor-α.

Article Snippet: The sections were incubated with rabbit anti-mouse PD-L1 polyclonal antibodies (dilution, 1:200; Abcam, Cambridge, MA, USA; cat. no. ab58810) overnight at 4°C, followed by incubation with biotinylated goat anti-rabbit secondary antibodies (1:5,000; ZSGB-BIO, Beijing, China) for 30 min at room temperature, and with streptavidin-peroxidase complex solution for another 30 min at room temperature.

Techniques: Expressing, Flow Cytometry, Cell Culture, Incubation

Journal: Molecular Medicine Reports

Article Title: Expression of programmed death 1 ligand 1 on periodontal tissue cells as a possible protective feedback mechanism against periodontal tissue destruction

doi: 10.3892/mmr.2016.4824

Figure Lengend Snippet: Expression of PD-L1 is correlated with the severity of periodontitis in the experimental periodontitis model. (A) Flow cytometry histograms of the expression of PD-L1 on the surface of periodontal tissue cells from healthy mice, and mice with mild periodontitis and severe periodontitis. (B) Expression of PD-L1 in periodontal tissues from the three groups, detected by immunohistochemical staining (magnification, ×400). (C) Comparison of the expression of PDL-1 in periodontal tissues between the three groups ( ‡ P<0.01, compared with the healthy control). (D) Comparison of the expression of PD-L1 on the surface of the periodontal tissue cells in the three groups ( * P<0.05, compared with the healthy control; † P<0.05, between the mild and severe periodontitis groups). (E) Comparison of the expression of PD-1 on the surface of the periodontal tissue cells between the three groups (F) Flow cytometry histograms of the expression of PD-1 on the surface of periodontal tissue cells from the three groups. No significant differences in the expression of PD-1 were observed between the three groups. Data are expressed as the mean ± standard error of the mean of three independent experiments. PD-L1, programmed death 1 ligand 1; APC, antigen-presenting cell.

Article Snippet: The sections were incubated with rabbit anti-mouse PD-L1 polyclonal antibodies (dilution, 1:200; Abcam, Cambridge, MA, USA; cat. no. ab58810) overnight at 4°C, followed by incubation with biotinylated goat anti-rabbit secondary antibodies (1:5,000; ZSGB-BIO, Beijing, China) for 30 min at room temperature, and with streptavidin-peroxidase complex solution for another 30 min at room temperature.

Techniques: Expressing, Flow Cytometry, Immunohistochemical staining, Staining