csf1 cytokine  (GenScript corporation)

Journal: Molecular oral microbiology

Article Title: Interleukin-34 permits Porphyromonas gingivalis survival and NF- κ B p65 inhibition in macrophages

doi: 10.1111/omi.12366

Figure Lengend Snippet: Interleukin-34 (IL-34) and colony-stimulating factor 1 (CSF1) equally differentiate blood monocytes into macrophages. Flow cytometry analysis of (a) CD14 and (b) CD163 on undifferentiated monocytes as well as CSF1 and IL-34 differentiated macrophages. The mean fluorescence intensity (MFI) of CD14 is not significantly different between all three cell types, while CD163 expression is increased significantly upon differentiation of macrophages in the presence of both IL-34 and CSF1. In all cases, ≥ 95% of differentiated macrophages were positive for CD14 and CD163. Shown is the average ± standard error of 4–9 independent experiments, * indicates p < 0.05 by one-way ANOVA with Dunnett’s multiple comparison

Article Snippet: Macrophages were generated from blood monocytes in six-well culture plates in culture medium (Roswell Park Memorial Institute (RPMI)) 1640 medium [Corning] containing 10% Fetal Bovine Serum (FBS) (Caisson Labs), 100 IU/ml penicillin, 100 μ g/ml streptomycin, and 0.25 μ g/ml amphotericin) for 6 days supplemented with 50 ng/ml CSF1 or 50 ng/ml IL-34 (GenScript; Foucher et al., 2013 ).

Techniques: Flow Cytometry, Fluorescence, Expressing, Comparison

Journal: Molecular oral microbiology

Article Title: Interleukin-34 permits Porphyromonas gingivalis survival and NF- κ B p65 inhibition in macrophages

doi: 10.1111/omi.12366

Figure Lengend Snippet: IL-34 macrophages are less efficient at killing internalized Porphyromonas gingivalis. (a) CSF1 and IL-34 macrophages are equally efficient at internalizing P. gingivalis. Shown are the average (± standard error) internalized bacteria per macrophage after 30 min co-incubation (MOI 10:1) of four independent experiments with bacteria quantified within a minimum of 100 macrophages per experiment. (b) Percent survival of internalized P. gingivalis within macrophages for 60 or 120 min post-uptake using an antibiotic protection assay. Shown is the mean (± standard error) of 3–4 independent experiments, p-value calculated with (a) an unpaired t-test and (b) a two-way ANOVA followed by Šídák’s multiple comparison test of survival of P. gingivalis within IL-34 and CSF1-differentiated macrophages at each timepoint

Article Snippet: Macrophages were generated from blood monocytes in six-well culture plates in culture medium (Roswell Park Memorial Institute (RPMI)) 1640 medium [Corning] containing 10% Fetal Bovine Serum (FBS) (Caisson Labs), 100 IU/ml penicillin, 100 μ g/ml streptomycin, and 0.25 μ g/ml amphotericin) for 6 days supplemented with 50 ng/ml CSF1 or 50 ng/ml IL-34 (GenScript; Foucher et al., 2013 ).

Techniques: Bacteria, Incubation, Comparison

Journal: Molecular oral microbiology

Article Title: Interleukin-34 permits Porphyromonas gingivalis survival and NF- κ B p65 inhibition in macrophages

doi: 10.1111/omi.12366

Figure Lengend Snippet: IL-34 downregulates intracellular reactive oxygen species (ROS) production from macrophages. Luminol-based ROS detection was used to quantify the ROS response by CSF1- and IL-34-differentiated macrophages during the activation with phorbol myristate acetate (100 nm). (a) Representative panel for the time course of luminol luminescence detection, shown in relative luminescence units (RLU). (b) Mean peak RLU (with standard error) was detected over three independent experiments. * p < 0.01 (paired t-test)

Article Snippet: Macrophages were generated from blood monocytes in six-well culture plates in culture medium (Roswell Park Memorial Institute (RPMI)) 1640 medium [Corning] containing 10% Fetal Bovine Serum (FBS) (Caisson Labs), 100 IU/ml penicillin, 100 μ g/ml streptomycin, and 0.25 μ g/ml amphotericin) for 6 days supplemented with 50 ng/ml CSF1 or 50 ng/ml IL-34 (GenScript; Foucher et al., 2013 ).

Techniques: Activation Assay

Journal: Molecular oral microbiology

Article Title: Interleukin-34 permits Porphyromonas gingivalis survival and NF- κ B p65 inhibition in macrophages

doi: 10.1111/omi.12366

Figure Lengend Snippet: Comparison of the surface expression of markers with known P. gingivalis interactions on IL-34 and CSF1 macrophages. The majority of markers with known interactions with P. gingivalis are not significantly different between CSF1- and IL-34-differentiated macrophages, with the exception of dendritic cell-specific intercellular adhesion molecule-grabbing nonintegrin (DC-SIGN). Macrophages matured in the presence of either CSF1 or IL-34 were labeled with CD14 and one or more of C5aR, CD11b, TLR 2, TLR 4, or DC-SIGN and analyzed by flow cytometry. The cells were gated for positive CD14 (≥ 90% of differentiated cells), and the MFI of the additional markers was determined. Representative histograms are shown, along with the mean (± standard error) of at least four independent experiments per marker tested. p-values were determined by paired (IL-34 vs. CSF1) t-tests.

Article Snippet: Macrophages were generated from blood monocytes in six-well culture plates in culture medium (Roswell Park Memorial Institute (RPMI)) 1640 medium [Corning] containing 10% Fetal Bovine Serum (FBS) (Caisson Labs), 100 IU/ml penicillin, 100 μ g/ml streptomycin, and 0.25 μ g/ml amphotericin) for 6 days supplemented with 50 ng/ml CSF1 or 50 ng/ml IL-34 (GenScript; Foucher et al., 2013 ).

Techniques: Comparison, Expressing, Labeling, Flow Cytometry, Marker

Journal: Molecular oral microbiology

Article Title: Interleukin-34 permits Porphyromonas gingivalis survival and NF- κ B p65 inhibition in macrophages

doi: 10.1111/omi.12366

Figure Lengend Snippet: IL-34-matured macrophages produce less IL-8 than CSF1-matured macrophages. IL-34- or CSF1-differentiated macrophages were co-incubated with P. gingivalis for 24 h, with media samples removed at 2, 6, and 24 h and subsequently analyzed by multiplex ELISA. The analytes that had significant release above control macrophages are shown. Four independent experiments were performed, mean ± standard error is shown. p-values were determined by two-way ANOVA followed by Šídák’s multiple comparison test of IL-34 versus CSF1-differentiated macrophages at each timepoint: those without p-value indicated have a p > 0.05. Significantly less IL-8 was produced by IL-34-differentiated macrophages than by CSF1-differentiated macrophages

Article Snippet: Macrophages were generated from blood monocytes in six-well culture plates in culture medium (Roswell Park Memorial Institute (RPMI)) 1640 medium [Corning] containing 10% Fetal Bovine Serum (FBS) (Caisson Labs), 100 IU/ml penicillin, 100 μ g/ml streptomycin, and 0.25 μ g/ml amphotericin) for 6 days supplemented with 50 ng/ml CSF1 or 50 ng/ml IL-34 (GenScript; Foucher et al., 2013 ).

Techniques: Incubation, Multiplex Assay, Enzyme-linked Immunosorbent Assay, Control, Comparison, Produced

Journal: Molecular oral microbiology

Article Title: Interleukin-34 permits Porphyromonas gingivalis survival and NF- κ B p65 inhibition in macrophages

doi: 10.1111/omi.12366

Figure Lengend Snippet: Dephosphorylation of NF-κB p65 is associated with lower levels of IL-8 in IL-34-differentiated macrophages. IL-34- or CSF1-differentiated macrophages were co-incubated with P. gingivalis for 2 h Total cell lysates were assessed by western blot using phospho-NF-κB p65 (Ser-536) or total NF-κB p65 antibodies. Anti-mouse actin was used as an internal control. Data shown in Panel B are representative of three independent experiments with similar results; Panel A shows the resulting densitometry analysis of the ratio between phosphorylated and total NF-κB. Statistical analysis (one-way ANOVA with pairwise comparison) indicates a significant difference in the phosphorylated NF-κB between IL-34- and CSF1-differentiated macrophages incubated with P. gingivalis but not control or LPS-incubated macrophages.

Article Snippet: Macrophages were generated from blood monocytes in six-well culture plates in culture medium (Roswell Park Memorial Institute (RPMI)) 1640 medium [Corning] containing 10% Fetal Bovine Serum (FBS) (Caisson Labs), 100 IU/ml penicillin, 100 μ g/ml streptomycin, and 0.25 μ g/ml amphotericin) for 6 days supplemented with 50 ng/ml CSF1 or 50 ng/ml IL-34 (GenScript; Foucher et al., 2013 ).

Techniques: De-Phosphorylation Assay, Incubation, Western Blot, Control, Comparison

Journal: Molecular oral microbiology

Article Title: Interleukin-34 permits Porphyromonas gingivalis survival and NF- κ B p65 inhibition in macrophages

doi: 10.1111/omi.12366

Figure Lengend Snippet: IL-34-matured macrophages produce less IL-8 upon NF-kB p65-specific inhibition. Human peripheral blood monocytes were differentiated with CSF1 (50 ng/ml) and IL-34 (50 ng/ml)for 6 days in 24-well tissue culture plates (2 × 105 cells/well). The differentiated macrophages were pre-incubated with NFkB p65 inhibitor (NBP2-29321, 100, μM) and control peptide (NBP2-29334, 100 μM) overnight. Cells were then stimulated with LPS (0.1 μg/ml) for 16 h. The cell supernatants were analyzed for the production of IL-8 by ELISA. Shown is the average ± standard error, **** indicates p < 0.0001 by one-way ANOVA with Dunnett’s multiple comparison test

Article Snippet: Macrophages were generated from blood monocytes in six-well culture plates in culture medium (Roswell Park Memorial Institute (RPMI)) 1640 medium [Corning] containing 10% Fetal Bovine Serum (FBS) (Caisson Labs), 100 IU/ml penicillin, 100 μ g/ml streptomycin, and 0.25 μ g/ml amphotericin) for 6 days supplemented with 50 ng/ml CSF1 or 50 ng/ml IL-34 (GenScript; Foucher et al., 2013 ).

Techniques: Inhibition, Incubation, Control, Enzyme-linked Immunosorbent Assay, Comparison