type p gingivalis strain atcc 33277  (ATCC)

Journal: Journal of Oral Microbiology

Article Title: Macrophage activation and invasion by P. gingivalis is modulated by PPAD and accessory fimbriae subunits

doi: 10.1080/20002297.2026.2638646

Figure Lengend Snippet: P. gingivalis strains with fimbriae depleted of all accessory subunits (DAP) poorly activate MDM pro-inflammatory responses. MDMs were treated for A) 4 h or B) 24 h with purified fimbriae (1 μg/ml) from various P. gingivalis strains, including ATCC 33277 WT (fimbriae WT) and ∆PPAD mutant (fimbriae ∆PPAD), as well as mutants of accessory fimbrial subunits, ∆ fimE mutant (fimbriae ∆FimE) and ∆ fimC mutant (fimbriae ∆FimC). A) Relative mRNA expression of PGE2 synthesis-related gene COX2 and cytokines IL6 and IL8 ; n = 4–7. B) Secretion of IL-6 and IL-8; n = 5–9. Data are presented as the mean ± SEM compared to WT fimbriae, with significance levels indicated as **** p < 0.0001; *** p < 0.001; ** p < 0.01; * p < 0.05.

Article Snippet: We thoroughly assessed how PPAD affects inflammatory and antibacterial responses of macrophages to the fimbriated wild-type P. gingivalis strain ATCC 33277.

Techniques: Purification, Mutagenesis, Expressing

Journal: Journal of Oral Microbiology

Article Title: Macrophage activation and invasion by P. gingivalis is modulated by PPAD and accessory fimbriae subunits

doi: 10.1080/20002297.2026.2638646

Figure Lengend Snippet: LPS contamination in fimbriae preparations is negligible and does not influence the pro-inflammatory activation of MDMs. A) MDMs were treated for 24 h with purified fimbriae (1 μg/ml) from ATCC 33277 WT (fimbriae WT) and ∆PPAD mutant (fimbriae ∆PPAD) strains, as well as LPS-free fimbriae from both strains; n = 3. B) hMDMs were preincubated for 30 min with control (IgG1) or TLR4-blocking antibody (anti-TLR4), then treated for 24 h with purified fimbriae (1 μg/ml) from ATCC 33277 WT (fimbriae WT) and the ∆ ppad mutant (fimbriae ∆PPAD) strains; n = 3. Levels of IL-6 and IL-8 secretion were subsequently measured. Data are presented as mean ± SEM. ‘ns’ indicates not statistically significant. Statistically significant differences are shown in A) between LPS-free fimbriae and fimbriae without the LPS removal step; in B) between IgG1 and anti-TLR4 pretreated samples.

Article Snippet: We thoroughly assessed how PPAD affects inflammatory and antibacterial responses of macrophages to the fimbriated wild-type P. gingivalis strain ATCC 33277.

Techniques: Activation Assay, Purification, Mutagenesis, Control, Blocking Assay

Journal: Journal of Oral Microbiology

Article Title: Macrophage activation and invasion by P. gingivalis is modulated by PPAD and accessory fimbriae subunits

doi: 10.1080/20002297.2026.2638646

Figure Lengend Snippet: PPAD and fimbriae mutants show differences in adhesion and safe entry to host cells. Flow cytometry analysis of MDMs infected for 30 min at MOI = 20 or 100 with P. gingivalis ATCC 33277 WT and PPAD mutant strains—including its isogenic total (∆PPAD) and catalytically inactive (C351A PPAD) strains, as well as fimbriae (∆FimA) mutants—labelled with A) CFSE or B) pHRODO-Red (pHRODO). Phagocytosis and adhesion rates were determined as the percentages of CFSE and pHRODO-Red positive cells, relative to the WT strain; n = 4. Results are shown as mean ± SEM. ** p < 0.01; * p < 0.05; ns, not significant. Comparisons are made to the ATCC WT strain. C) Widefield fluorescence microscopy of MDMs infected for 30 min at MOI 100 with CFSE-labelled P. gingivalis ATCC WT, ∆PPAD, C351A PPAD, ∆FimA, ∆FimE ∆FimC, and W83 strains. Nuclei were stained with DAPI, and actin was stained with phalloidin conjugated to Alexa Fluor 647. Scale bars: 30 µm. Images are representative of three independent experiments. White arrows indicate bacteria that are not internalised.

Article Snippet: We thoroughly assessed how PPAD affects inflammatory and antibacterial responses of macrophages to the fimbriated wild-type P. gingivalis strain ATCC 33277.

Techniques: Flow Cytometry, Infection, Mutagenesis, Fluorescence, Microscopy, Staining, Bacteria

Journal: Journal of Oral Microbiology

Article Title: Macrophage activation and invasion by P. gingivalis is modulated by PPAD and accessory fimbriae subunits

doi: 10.1080/20002297.2026.2638646

Figure Lengend Snippet: Phosphorylation of Akt is enhanced by PPAD and accessory fimbriae subunits, with the PI3K/Akt pathway being essential for IL-6 but not for IL-8 secretion. MDMs were infected for A) 10 min, 30 min, or 2 h with P. gingivalis ATCC 33277 WT strain (ATCC WT) and its isogenic PPAD mutant (∆PPAD), or B) for 30 min with ATCC 33277-derived PPAD: total (∆PPAD) and catalytically inactive (C351A PPAD), fimbriae: main FimA subunit (∆ fimA ), accessory subunits FimE (∆ fimE ), and FimC (∆ fimC ) mutants, or the W83 WT strain at MOI 20. Akt phosphorylation levels were determined by western blot analysis, with total Akt as a control and β -actin as the loading control. Representative blots are shown, and densitometry results ( n = 3) are presented as mean ± SEM. *** p < 0.001; ** p < 0.01; * p < 0.05; ns, not significant. In B), data are compared with the ATCC WT strain. C) MDMs were pretreated for 30 min with an Akt inhibitor or left untreated, then infected for 24 h at MOI 20 with P. gingivalis ATCC WT, ∆PPAD, C351A PPAD, ∆FimA, ∆FimE, ∆FimC, and W83 strains. Secretion of IL-6 and IL-8 was measured by ELISA; n = 4−7. Results are shown as mean ± SEM; **** p < 0.0001; *** p < 0.001; ** p < 0.01; * p < 0.05 compared to ATCC WT without Akt inhibitor; ns, not significant.

Article Snippet: We thoroughly assessed how PPAD affects inflammatory and antibacterial responses of macrophages to the fimbriated wild-type P. gingivalis strain ATCC 33277.

Techniques: Phospho-proteomics, Infection, Mutagenesis, Derivative Assay, Western Blot, Control, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Dual species interactions shield Campylobacter against multiple antibiotics

doi: 10.64898/2026.02.27.708565

Figure Lengend Snippet: Resistance patterns of MC14-02991.B and derivatives Cj14-02991.8 and Efs22-05285.1. MIC values above the ECOFF for mixed samples and purified strains are marked bold and the respective isolates are categorized as non-wildtype for this antimicrobial. If a MIC value is below the ECOFF, the corresponding isolate is categorized as wildtype for this antimicrobial. Since there is no C. jejuni ECOFF for meropenem available from EUCAST, ertapenem ECOFF is given instead. Currently, no Enterococcus ECOFFS or breakpoints are available for azithromycin, clindamycin and nourseothricin (EUCAST) (see Material and Methods) and therefore the respective MIC values cannot be categorized into wildtype/ non-wildtype. The ASA cluster is underlined. See material and methods for full panel.

Article Snippet: To assess whether this is a more general feature of C. jejuni / E. faecium interaction, we applied this assay on the type strain C. jejuni ATCC 33560 with and without co-incubation with MDR E. faecium strain EFm22-05284.1.

Techniques: Purification

Journal: bioRxiv

Article Title: Dual species interactions shield Campylobacter against multiple antibiotics

doi: 10.64898/2026.02.27.708565

Figure Lengend Snippet: : Images from two colonies each of mixed samples MC14-02991.B and MC20-00984.21 are shown. Bacteria were grown on Brucella agar or CCDA. For sample MC14-02991.B, coccoid bacteria were found in all investigated colonies of the mixed cultures. The cocci were either intermingled with Campylobacter , as shown here, or present in smaller groups. For MC20-00984.21, coccoid bacteria were found in some but not in all investigated colonies of the mixed samples. They were always intermingled with the spiral shaped Campylobacter and never appeared in groups. : Images from colonies of separated strains. From left to right: C. jejuni (Cj14-02991.8), E. faecalis (Efs22-05285.1), C. jejuni (Cj-20-00984.4) and E. faecium (Efm22-05284.1). Bacteria were grown on Brucella Agar or CCDA. On Brucella agar, C. jejuni may develop an unusual round to pleomorphic morphology (arrow) which can be the dominant form (see also , Cj ATCC 33560). : Images from colonies of reference isolates. From left to right: C. jejuni (Cj ATCC 33560) grown on CCDA, C. jejuni (Cj ATCC 33560) grown on Brucella agar, E. faecalis (Efs ATCC 29212) grown on Brucella agar, E. faecium (Efm ATCC 19434) grown on CCDA.

Article Snippet: To assess whether this is a more general feature of C. jejuni / E. faecium interaction, we applied this assay on the type strain C. jejuni ATCC 33560 with and without co-incubation with MDR E. faecium strain EFm22-05284.1.

Techniques: Bacteria

Journal: bioRxiv

Article Title: Dual species interactions shield Campylobacter against multiple antibiotics

doi: 10.64898/2026.02.27.708565

Figure Lengend Snippet: First a microdilution assay was performed for every test antimicrobial for 24h incubation at 42°C (microaerophilic). For re-isolation of C. jejuni , 5 µl suspension from each well was transferred to Brucella agar containing Campylobacter selective supplement and incubated at 42°C for 48h under microaerophilic conditions. The agar did not allow growth of E. faecalis or sufficient growth of E. faecium within the given time of 48h. The isolation MIC is the lowest concentration of the antimicrobial, where no C. jejuni could be isolated from the wells, i.e. no growth was visible on the agar plate. The test was used for the following antimicrobials: azithromycin, clindamycin, tetracycline, gentamicin, meropenem and nourseothricin.

Article Snippet: To assess whether this is a more general feature of C. jejuni / E. faecium interaction, we applied this assay on the type strain C. jejuni ATCC 33560 with and without co-incubation with MDR E. faecium strain EFm22-05284.1.

Techniques: Microdilution Assay, Incubation, Isolation, Suspension, Concentration Assay