Journal: Innate Immunity

Article Title: Susceptibility of primary human airway epithelial cells to Bordetella pertussis adenylate cyclase toxin in two- and three-dimensional culture conditions

doi: 10.1177/1753425920979354

Figure Lengend Snippet: Airway epithelial cells do not express CD11b. Expression of CD11b in airway epithelial cells and THP-1 was analysed by flow cytometry. CD11b-positive cells were not found in HNEC (a), HTEC (b) or HBEC3-KT (c) samples. THP-1 populations were CD11b-positive (d). Blue histograms show isotype control-stained cells and red histograms show cells stained with an anti-CD11b Ab. The figure shows the histograms of one representative experiment ( n =3).

Article Snippet: 14, Airway epithelial cells were grown in Airway Epithelial Cell Growth Medium (AECG; #PB-C-MH-350-0099, PeloBiotech, Martinsried, Germany).

Techniques: Expressing, Flow Cytometry, Control, Staining

Journal: Innate Immunity

Article Title: Susceptibility of primary human airway epithelial cells to Bordetella pertussis adenylate cyclase toxin in two- and three-dimensional culture conditions

doi: 10.1177/1753425920979354

Figure Lengend Snippet: CyaA affects intracellular cAMP concentration and IL-6 secretion in human airway epithelial cells. (a) HNEC, HTEC and HBEC3-KT were incubated with CyaA or CyaA-AC – and intracellular cAMP concentration was analysed ( n = 4). (b) Quantification of human IL-6 secreted by HNEC, HTEC and HBEC3-KT populations after treatment with CyaA or CyaA-AC – was assayed by ELISA of the supernatants ( n = 3). (c) Quantification of human IL-6 secreted by human nasal mucosa tissue models after treatment with CyaA or CyaA-AC – ( n =3). All data are presented as means (bars) + SEM (error bars). Differences after CyaA incubation compared to the control group (TUC) are indicated (* P ≤ 0.05, $ P > 0.05 ≤ 0.1, ANOVA). Statistically significant differences of IL-6 basal levels are indicated ( # P < 0.05, ANOVA). Significant cell type-specific differences are indicated ( & P < 0.05, ANOVA).

Article Snippet: 14, Airway epithelial cells were grown in Airway Epithelial Cell Growth Medium (AECG; #PB-C-MH-350-0099, PeloBiotech, Martinsried, Germany).

Techniques: Concentration Assay, Incubation, Enzyme-linked Immunosorbent Assay, Control

Journal:

Article Title: Porphyromonas gingivalis Gingipains and Adhesion to Epithelial Cells

doi: 10.1128/IAI.69.5.3048-3056.2001

Figure Lengend Snippet: Effects of Arg-gingipain mutations on P. gingivalis attachment to KB oral epithelial cells

Article Snippet: However, the sonicate did not contain Rgp or Kgp proteolytic activity (data not shown), and although we have not ruled out the possibility that a catalytically inactive Kgp precursor was present in this strain, it is also possible that adhesin peptides were derived from cell-associated HagA. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 5 caption a7 Western blot analysis of epithelial cell extracts showing the capture of proteins from cell sonicates of ATCC 33277 and KDP112.

Techniques: Activity Assay, Bacteria

Journal:

Article Title: Porphyromonas gingivalis Gingipains and Adhesion to Epithelial Cells

doi: 10.1128/IAI.69.5.3048-3056.2001

Figure Lengend Snippet: Western blot of KB epithelial cell extracts after capture of adhesins from VDS fractions of P. gingivalis parent and rgp mutant strains. Before incubation with KB cells, VDS fractions were untreated (−), treated with 500 μM TLCK (T), or heated at 100°C for 5 min (triangles). (A) Blots were probed with antibody to gingipain adhesin domains, and cross-reacting adhesin peptides are indicated by arrowheads. (B) Blots were probed with fimbrillin antibody. Fimbrillin (indicated by the arrowhead) showed more intense reactivity in boiled VDS fractions. WT, wild type.

Article Snippet: However, the sonicate did not contain Rgp or Kgp proteolytic activity (data not shown), and although we have not ruled out the possibility that a catalytically inactive Kgp precursor was present in this strain, it is also possible that adhesin peptides were derived from cell-associated HagA. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 5 caption a7 Western blot analysis of epithelial cell extracts showing the capture of proteins from cell sonicates of ATCC 33277 and KDP112.

Techniques: Western Blot, Mutagenesis, Incubation

Journal:

Article Title: Porphyromonas gingivalis Gingipains and Adhesion to Epithelial Cells

doi: 10.1128/IAI.69.5.3048-3056.2001

Figure Lengend Snippet: Western blot analysis of epithelial cell extracts showing the capture of proteins from cell sonicates of ATCC 33277 and KDP112. Blots were probed with antiadhesin antibody. Triangles denote heat-treated sonicates. The 44-, 39-, 27-, and 19-kDa adhesin peptides (indicated by arrowheads) were present in the cell sonicates of KDP112 and ATCC 33277.

Article Snippet: However, the sonicate did not contain Rgp or Kgp proteolytic activity (data not shown), and although we have not ruled out the possibility that a catalytically inactive Kgp precursor was present in this strain, it is also possible that adhesin peptides were derived from cell-associated HagA. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 5 caption a7 Western blot analysis of epithelial cell extracts showing the capture of proteins from cell sonicates of ATCC 33277 and KDP112.

Techniques: Western Blot

Journal:

Article Title: Porphyromonas gingivalis Gingipains and Adhesion to Epithelial Cells

doi: 10.1128/IAI.69.5.3048-3056.2001

Figure Lengend Snippet: Gingipain-mediated adhesion and detachment of P. gingivalis to epithelial cells. We hypothesize that adhesion is mediated by gingipain adhesin peptides localized at the surface of the outer membrane and in membrane vesicles. Rgp catalytic activities, either noncovalently linked with adhesin domains or as soluble proteins, modulate adhesion through digestion of binding substrate. With the parent strain, ATCC 33277, the monolayer adhesion assay measures the net reaction of bacterial attachment to and detachment from receptors or extracellular matrix proteins at the epithelial cell surface. When catalytic activity is reduced by mutation (KDP112), the assay measures the large number of bacteria that attach (via Kgp or HagA adhesin peptides) and accumulate on the monolayers but do not detach or do so very slowly.

Article Snippet: However, the sonicate did not contain Rgp or Kgp proteolytic activity (data not shown), and although we have not ruled out the possibility that a catalytically inactive Kgp precursor was present in this strain, it is also possible that adhesin peptides were derived from cell-associated HagA. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 5 caption a7 Western blot analysis of epithelial cell extracts showing the capture of proteins from cell sonicates of ATCC 33277 and KDP112.

Techniques: Membrane, Binding Assay, Cell Adhesion Assay, Activity Assay, Mutagenesis, Bacteria