wild type strains  (DSMZ)

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: Molecular Therapy Oncology

Article Title: Improved oncolytic and immunostimulatory activity of the spontaneous jin-3 reovirus mutant in preclinical bladder cancer models

doi: 10.1016/j.omton.2026.201128

Figure Lengend Snippet: Comparison of the oncolytic effects of mutant jin-3 versus R124 reovirus in bladder cancer cell lines in vitro (A) Mean viral S4Q copy number (log fold change S4Q mRNA expression vs. mock treated cells) in bladder cancer cell lines exposed to a range of MOI (0.01–0.1–1–10) of either R124 or jin-3 reoviruses after 24 h. In red, the highest infection with virus is shown. Two-way ANOVA followed by Tukey’s post hoc test. n = 3 (2 replicates). UM-UC-3 cells R124 reovirus MOI1 vs. mock p = 0.0184; ∗∗∗ p < .001, $$$ p < .001, asterisks indicate mock versus reovirus infection, and dollar signs indicate R124 versus jin-3. (B) Viral load (% of viable, single cells expressing Sigma-3 protein) in bladder cancer cell lines exposed to MOI10 of either R124 or jin-3 after 72 h mean (SD) ( N = 3). Two-way ANOVA followed by Tukey’s post hoc test. ∗∗∗ p < .001, $$$ p < .001, asterisks indicate mock versus reovirus infection, and dollar signs indicate R124 versus jin-3. (C) Confocal images of viral protein (Sigma-3, green) and DAPI (blue)-stained bladder cancer cells treated with MOI10 of R124 or jin-3 for 72h. Scale bars, 25 μm. (D) Mean percentage of viable cells after exposure to a range of MOI of either R124 or jin-3 for 6 days. n = 3 (6 replicates). p values are depicted (vs. mock; when 2 depicted upper p value is R124 vs. jin-3 ); ∗∗∗ p < .001, $$$ p < .001, asterisks indicate mock versus reovirus infection, and dollar signs indicate R124 versus jin-3. (E) Percentage of single, viable, JAM-A protein expressing cells. Mean (SD), n = 3. MOI = multiplicity of infection.

Article Snippet: Wild-type T3D reovirus strain R124 was plaque-purified from the wild-type reovirus T3D (ATCC) on HER911 cells., Reovirus mutant jin-3 was isolated from JAM-A-deficient U118MG cells after passaging of the wild-type T3D strain R124.

Techniques: Comparison, Mutagenesis, In Vitro, Expressing, Infection, Virus, Staining

Journal: Molecular Therapy Oncology

Article Title: Improved oncolytic and immunostimulatory activity of the spontaneous jin-3 reovirus mutant in preclinical bladder cancer models

doi: 10.1016/j.omton.2026.201128

Figure Lengend Snippet: Comparison of the oncolytic effects of reovirus mutant jin-3 versus wild-type reovirus R124 in 3D cultures in vitro TM00024 bladder PDXOs were exposed to the indicated plaque-forming units (PFUs) of reoviruses for 3 or 7 days. (A) Mean (SD) viral copy number of TM00024 PDXOs treated for 3 days with the indicated PFUs of the reoviruses. Two-way ANOVA followed by Tukey’s post hoc comparison ( n = 3) ∗∗∗ p < .001. (B–F) Viability (mean [SD] was measured using cell titer glo 3D after days 3 and 7 of reovirus exposure. N = 3 (3 replicates). Two-way ANOVA followed by Tukey’s post hoc comparison. ∗∗∗ p < .001. (C) Confocal images of PDXOs (day 7) stained for respectively H&E, panKRT (red); Sigma-3, c-CASP3, or PCNA (green); and DAPI (blue). Scale bars, 20 μm. Cells expressing the respective proteins Sigma-3 (D), c-CASP-3 (E), and nuclear PCNA (F) were counted with ImageJ and divided by the amount of panKRT+_DAPI+ cells. At least five fields were scored per technical replicate. Mean (SD) of N = 3 (3 replicates). ∗∗∗ p < .001, asterisks indicate reovirus infection versus mock same day. Two-way ANOVA followed by Tukey’s post hoc comparison.

Article Snippet: Wild-type T3D reovirus strain R124 was plaque-purified from the wild-type reovirus T3D (ATCC) on HER911 cells., Reovirus mutant jin-3 was isolated from JAM-A-deficient U118MG cells after passaging of the wild-type T3D strain R124.

Techniques: Comparison, Mutagenesis, In Vitro, Staining, Expressing, Infection

Journal: Molecular Therapy Oncology

Article Title: Improved oncolytic and immunostimulatory activity of the spontaneous jin-3 reovirus mutant in preclinical bladder cancer models

doi: 10.1016/j.omton.2026.201128

Figure Lengend Snippet: Comparison of jin-3 and R124 reovirus infection in ex vivo cultured tumor tissue Explanted tissue slices from either RT-112 CDX (A–E) or TM00024 PDX (F–J) were exposed to the indicated PFU of R124 or jin-3 reovirus for 3 days. Tissues were stained for H&E, panKRT (red); Sigma-3, c-CASP3 or PCNA (green), type I collagen (white), and DAPI (blue). Representative confocal images are shown. Scale bars, 20 μm. Cells expressing the respective proteins were counted with ImageJ and divided by the number of panKRT+_DAPI+ cells. At least four fields were scored per technical replicate. Mean (SD) of N = 2 (4 replicates). The ratio fragmented tumor cells was measured by the number of fragmented cells/total tumor cells (E and J). ∗∗∗ p < .001, reovirus infection versus mock. Mean (SD), n = 2 (4 replicates). Two-way ANOVA followed by Tukey’s post hoc comparison.

Article Snippet: Wild-type T3D reovirus strain R124 was plaque-purified from the wild-type reovirus T3D (ATCC) on HER911 cells., Reovirus mutant jin-3 was isolated from JAM-A-deficient U118MG cells after passaging of the wild-type T3D strain R124.

Techniques: Comparison, Infection, Ex Vivo, Cell Culture, Staining, Expressing

Journal: Molecular Therapy Oncology

Article Title: Improved oncolytic and immunostimulatory activity of the spontaneous jin-3 reovirus mutant in preclinical bladder cancer models

doi: 10.1016/j.omton.2026.201128

Figure Lengend Snippet: Comparison of jin-3 and R124 reovirus infection in ex vivo cultured tumor tissue slices from human bladder cancer patients Explanted tissue slices from patients ( n = 15) diagnosed with bladder cancer were exposed to either mock, R124, or jin-3 reovirus for 3 days and stained for viral protein, c-CASP-3, PCNA (proliferation), and integrity of the cell (nuclear DAPI and panKRT tumor marker). (A–E) Representative confocal images of the mock and 10 7 PFU condition of both viruses from five patients ranging from low-risk NMIBC to MIBC (viral protein [green, Sigma-3], tumor markers [red, panKRT], and nuclei [DAPI, blue]. Scale bars, 20 μm). Cells expressing the respective proteins [(B) Sigma-3, (C) c-CASP3, or (D) nuclear PCNA] were counted with ImageJ and divided by the number of panKRT+_DAPI+ cells. At least four fields were scored per technical replicate. Mean (SD) (3–4 replicates) with each dot representing one bladder cancer patient. The ratio of fragmented tumor cells was measured by the number of fragmented cells/total tumor cells (E). ∗∗∗ p < 0.001, reovirus infection versus mock, one-way ANOVA followed by Tukey’ post hoc comparison.

Article Snippet: Wild-type T3D reovirus strain R124 was plaque-purified from the wild-type reovirus T3D (ATCC) on HER911 cells., Reovirus mutant jin-3 was isolated from JAM-A-deficient U118MG cells after passaging of the wild-type T3D strain R124.

Techniques: Comparison, Infection, Ex Vivo, Cell Culture, Staining, Marker, Expressing

Journal: Molecular Therapy Oncology

Article Title: Improved oncolytic and immunostimulatory activity of the spontaneous jin-3 reovirus mutant in preclinical bladder cancer models

doi: 10.1016/j.omton.2026.201128

Figure Lengend Snippet: Comparison of immune modulation induced by jin-3 and R124 reovirus treatment of bladder cancer cell lines Heat maps of various inflammatory cytokines and interferon-stimulated genes (log fold change mRNA expression [2 −ΔΔCt ] vs. mock treated cells) in bladder cancer cell lines UM-UC-3 (A), T24 (B), HT-1197 (C), RT-112 (D), RT-4 (E), TCCSUP (F), J82 (G), and PDXOs (H) exposed to a range of either R124 or jin-3 reoviruses after respectively 24 h (cell lines) and 3 days (PDXOs). p values are depicted (vs. mock; when 2 depicted upper p value is R124 vs. jin-3 ) ∗∗∗ p < .001, $$$ p < .001, asterisks indicate mock versus reovirus infection, and dollar signs indicate R124 versus jin-3.

Article Snippet: Wild-type T3D reovirus strain R124 was plaque-purified from the wild-type reovirus T3D (ATCC) on HER911 cells., Reovirus mutant jin-3 was isolated from JAM-A-deficient U118MG cells after passaging of the wild-type T3D strain R124.

Techniques: Comparison, Expressing, Infection

Journal: Molecular Therapy Oncology

Article Title: Improved oncolytic and immunostimulatory activity of the spontaneous jin-3 reovirus mutant in preclinical bladder cancer models

doi: 10.1016/j.omton.2026.201128

Figure Lengend Snippet: Comparison of immunogenic cell death induced by jin-3 and R124 reovirus treatment of bladder cancer cell lines (A–C) Cell lines were treated with oncolytic viruses for 48 h at an MOI of 10, and the DAMPs ecto-calreticulin (A) and ecto-HSP90 (B) were measured using flow cytometry. Secreted HMGB1 protein was measured with an ELISA after 48 h of treatment with a dose range of oncolytic viruses (C). ∗∗∗ p < 0.001, $$$ p < 0.001, asterisks indicate reovirus infection versus mock same day, and dollar signs indicate R124 vs. jin-3. Mean (SD), N = 3 (2 replicates). Two-way ANOVA followed by Tukey’s posthoc comparison. (D) Correlation graph of the HMGB1 release ( x axis), percentage of viable cells (size of the dots), and fold change of ecto-CRT ( y axis)-positive cells at MOI 10 of the indicated virus.

Article Snippet: Wild-type T3D reovirus strain R124 was plaque-purified from the wild-type reovirus T3D (ATCC) on HER911 cells., Reovirus mutant jin-3 was isolated from JAM-A-deficient U118MG cells after passaging of the wild-type T3D strain R124.

Techniques: Comparison, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Infection, Virus

Journal: Molecular Therapy Oncology

Article Title: Improved oncolytic and immunostimulatory activity of the spontaneous jin-3 reovirus mutant in preclinical bladder cancer models

doi: 10.1016/j.omton.2026.201128

Figure Lengend Snippet: Activation of PBMCs in RT-112 co-cultures (A) Brightfield images of RT-112 bladder cancer cells that were cultured in 60% Matrigel and allowed to form 3D structures for approximately 3 days. Subsequently, PBMCs were added at an effector: target ratio of 20:1 in absence or presence of either R124 or jin-3 reovirus at the indicated PFU for an additional 3 days. (B) Fragmented KRT18 was determined as an outcome measure for tumor cell killing 3 days after OV exposure. (C–E) CXCL10 levels and (D) IFN-γ levels were measured 3 days after OV exposure. ( n = 3, 2 replicates). Mean (SD). Two-way ANOVA with Tukey’s post hoc comparison. (E) Heatmap of various inflammatory cytokines and interferon-stimulated genes (log fold change mRNA expression [2 −ΔΔCt ] vs. mock treated cells) in 3D cultured RT-112 cells or RT-112 cells co-cultured with PBMCs exposed to either R124 or jin-3 reoviruses. Mean (SD). p values are depicted (vs. mock; when two depicted upper p value is R124 vs. jin-3 ) ∗∗∗ p < .001, $$$ p < .001, asterisks indicate mock versus reovirus infection and dollar signs R124 versus jin-3. , n = 2 (2 replicates). Two-way ANOVA followed by Tukey’s post hoc comparison.

Article Snippet: Wild-type T3D reovirus strain R124 was plaque-purified from the wild-type reovirus T3D (ATCC) on HER911 cells., Reovirus mutant jin-3 was isolated from JAM-A-deficient U118MG cells after passaging of the wild-type T3D strain R124.

Techniques: Activation Assay, Cell Culture, Comparison, Expressing, Infection

Journal: bioRxiv

Article Title: Synthetic Genome Shuffling of Poxviruses through Yeast for Next-Generation Oncolytic Platforms

doi: 10.64898/2026.03.06.710085

Figure Lengend Snippet: Yeast cells were co-transformed with fragmented pYCC3-VACV plasmid DNA together with CPXV and RPXV genomic DNA. Each resulting yeast colony is expected to contain a distinct chimeric poxvirus sequence (pYCC3-cPOX) generated through homologous recombination. DNA extracted from individual colonies was subsequently transfected into human cells pre-infected with the helper virus MVA, enabling the production of chimeric infectious viral particles.

Article Snippet: Wild-type Vaccinia virus strain Copenhagen (VACV) was obtained from Institut Mérieux (France), while wild-type Cowpox virus strain Brighton (CPXV; ATCC VR-302) and wild-type Rabbitpox virus strain Utrecht (RPXV; ATCC VR-1591) were obtained from ATCC (USA).

Techniques: Transformation Assay, Plasmid Preparation, Sequencing, Generated, Homologous Recombination, Transfection, Infection, Virus

Journal: bioRxiv

Article Title: Synthetic Genome Shuffling of Poxviruses through Yeast for Next-Generation Oncolytic Platforms

doi: 10.64898/2026.03.06.710085

Figure Lengend Snippet: (A) Brightfield microscopy images of plaque assays performed on HeLa cells infected with parental viruses (VACV, CPXV, RPXV) and with chimeric viruses rescued from TAR-shuffling clones (cPOX01-02, cPOX02-03, cPOX04-12, cPOX05-14, and cPOX06-18). HeLa cells were seeded at 5 × 10⁵ cells per well in 6-well plates and infected at a multiplicity of infection (MOI) of 10⁻⁵. Images were acquired 48 h post-infection at 4× magnification. (B) Representative comet-shaped plaques observed in A549 cells. A549 monolayers were infected with the indicated viruses, incubated for 48 h, and stained with crystal violet to visualize comet formation, reflecting enhanced viral spread

Article Snippet: Wild-type Vaccinia virus strain Copenhagen (VACV) was obtained from Institut Mérieux (France), while wild-type Cowpox virus strain Brighton (CPXV; ATCC VR-302) and wild-type Rabbitpox virus strain Utrecht (RPXV; ATCC VR-1591) were obtained from ATCC (USA).

Techniques: Microscopy, Infection, Clone Assay, Incubation, Staining

Journal: bioRxiv

Article Title: Synthetic Genome Shuffling of Poxviruses through Yeast for Next-Generation Oncolytic Platforms

doi: 10.64898/2026.03.06.710085

Figure Lengend Snippet: Yeast cells were co-transformed with fragmented pYCC3-VACV plasmid DNA together with CPXV and RPXV genomic DNA. Each resulting yeast colony is expected to contain a distinct chimeric poxvirus sequence (pYCC3-cPOX) generated through homologous recombination. DNA extracted from individual colonies was subsequently transfected into human cells pre-infected with the helper virus MVA, enabling the production of chimeric infectious viral particles.

Article Snippet: Wild-type Vaccinia virus strain Copenhagen (VACV) was obtained from Institut Mérieux (France), while wild-type Cowpox virus strain Brighton (CPXV; ATCC VR-302) and wild-type Rabbitpox virus strain Utrecht (RPXV; ATCC VR-1591) were obtained from ATCC (USA).

Techniques: Transformation Assay, Plasmid Preparation, Sequencing, Generated, Homologous Recombination, Transfection, Infection, Virus

Journal: bioRxiv

Article Title: Synthetic Genome Shuffling of Poxviruses through Yeast for Next-Generation Oncolytic Platforms

doi: 10.64898/2026.03.06.710085

Figure Lengend Snippet: (A) Brightfield microscopy images of plaque assays performed on HeLa cells infected with parental viruses (VACV, CPXV, RPXV) and with chimeric viruses rescued from TAR-shuffling clones (cPOX01-02, cPOX02-03, cPOX04-12, cPOX05-14, and cPOX06-18). HeLa cells were seeded at 5 × 10⁵ cells per well in 6-well plates and infected at a multiplicity of infection (MOI) of 10⁻⁵. Images were acquired 48 h post-infection at 4× magnification. (B) Representative comet-shaped plaques observed in A549 cells. A549 monolayers were infected with the indicated viruses, incubated for 48 h, and stained with crystal violet to visualize comet formation, reflecting enhanced viral spread

Article Snippet: Wild-type Vaccinia virus strain Copenhagen (VACV) was obtained from Institut Mérieux (France), while wild-type Cowpox virus strain Brighton (CPXV; ATCC VR-302) and wild-type Rabbitpox virus strain Utrecht (RPXV; ATCC VR-1591) were obtained from ATCC (USA).

Techniques: Microscopy, Infection, Clone Assay, Incubation, Staining

Journal: mBio

Article Title: Ligand binding drives proteolysis of the SmcR master transcription factor and controls quorum sensing-state transitions in Vibrio species

doi: 10.1128/mbio.03445-25

Figure Lengend Snippet: SmcR degradation controls the HCD to LCD transition. ( A–B ) Bioluminescence assays are plotted (Lux/OD 600 ) for V. vulnificus strains ATCC 27562 wild-type (WT) and isogenic mutants. Strains were treated with 25 µM PTSP (+) or an equivalent volume of DMSO solvent (−). Figures are representative of triplicate biological assays (shown in ).

Article Snippet: Isogenic strains containing mutations in smcR were generated by mutating wild-type V. vulnificus strain ATCC 27562.

Techniques: Solvent