hp strain 43504  (ATCC)

Journal: Molecular Medicine

Article Title: Helicobacter pylori VacA modulates TRAF1-mediated 4-1BB/NF-kappaB axis to induce host apoptosis and chronic inflammatory damage

doi: 10.1186/s10020-025-01349-5

Figure Lengend Snippet: Effects of VacA on TRAF1/4-1BB/NF-κB pathway/chemokine IL-8 axis- and apoptosis-related protein expression in GES-1 cells. a-d GES-1 cells infected with vacA + Hp strain and Δ vacA Hp strain at different MOIs (0, 10, 20, 50, 100, and 200) for 24 h or different time points (0 h, 6 h, 12 h, 24 h, and 36 h), MOI = 100. a, b Western blot analysis of the target proteins (phospho-p65, p65, TRAF1, 4-1BB, apoptosis-related protein Bcl-xl and Bax) levels. GAPDH was used as an internal reference. c, d the secretion levels of IL-8 from GES-1 cells after infection were measured by ELISA. Comparison between vacA + Hp strain-infected group and Δ vacA Hp strain-infected group under identical MOI conditions (c) or identical intervention time (d) showed statistical significance ( p < 0.001 for all comparisons). e Western blot analysis of the target proteins levels in GES-1 cells incubated with recombinant VacA protein at low concentrations (5 µg/ml and 10 µg/ml) for 48 h or at high concentrations (65 µg/ml) for 12 h. In addition, GES-1 cells incubated with isovolumetric protein buffer and cell culture medium served as a buffer control group and an untreated normal group, respectively. The relative protein expression level of TRAF1 and 4-1BB were compared between recombinant VacA protein-incubated group (10 µg/mL) and the correspondind buffer-incubated group, with statistically significant differences ( p = 0.041, p = 0.037, respectively). The relative protein expression level of p-p65, TRAF1, 4-1BB, Bcl-xl and Bax were compared between recombinant VacA protein-incubated group (65 µg/mL) and the correspondind buffer-incubated group, with statistically significant differences ( p < 0.001, p = 0.039, p = 0.044, p = 0.049, p = 0.047, respectively). f Western blot analysis of VacA, phospho-p65, p65, TRAF1, 4-1BB and apoptosis-related protein (Bcl-xl, Bax) levels in GES-1 cells transfected with pDsRED2-N1-HA/VacA for 48 h. g qRT‒PCR analysis of VacA, TRAF1, 4-1BB and IL-8 levels in GES-1 cells transfected with pDsRED2-N1-HA/VacA for 48 h. GAPDH was used as an internal control. Comparison of relative expression levels of VacA, TRAF1, 4-1BB, and IL-8 between the pDsRED2-N1-HA-VacA-transfected groups and the empty vector pDsRED2-N1-HA-transfected groups showed statistically significant differences ( p < 0.001, p = 0.007, p = 0.009, p = 0.005, respectively). h, i The secretion level of IL-8 in GES-1 cells after incubation with recombinant VacA protein ( h ) or transfection with pDsRED2-N1-HA/VacA ( i ) was measured by ELISA. h Levels of IL-8 were compared between recombinant VacA protein-incubated groups (at concentrations of 5, 10, and 65 µg/mL) and the correspondind buffer-incubated groups, with statistically significant differences ( p = 0.007, p < 0.001, p = 0.005, respectively). i Levels of IL-8 were compared between the pDsRED2-N1-HA-VacA-transfected groups (for 48 h and 72 h) and the empty vector pDsRED2-N1-HA-transfected groups, with statistically significant differences ( p = 0.039 and p = 0.0071). The figure presents the average of three independent experiments ( n = 3). Note: Comparisons were made between two groups using standard two-tailed Student’s t-test. The figure presents the average of three independent experiments ( n = 3). Data are presented as mean ± SEM. Error bars represent standard error of mean. *: p < 0.05. **: p < 0.01. ***: p < 0.001

Article Snippet: Subsequently, the mice were challenged using orogastric infusions of 300 μl of sterile Brucella broth, 5 × 10 9 CFU/ml vacA + Hp strain (the wild-type strain Hp ATCC 26695) or 5 × 10 9 CFU/ml ΔvacA Hp strain ( vacA -KO mutant Hp ATCC 26695) once every 2 days for a total of 9 infusions.

Techniques: Expressing, Infection, Western Blot, Enzyme-linked Immunosorbent Assay, Comparison, Incubation, Recombinant, Cell Culture, Control, Transfection, Plasmid Preparation, Two Tailed Test

Journal: Molecular Medicine

Article Title: Helicobacter pylori VacA modulates TRAF1-mediated 4-1BB/NF-kappaB axis to induce host apoptosis and chronic inflammatory damage

doi: 10.1186/s10020-025-01349-5

Figure Lengend Snippet: VacA activates the NF-κB pathway in GES-1 cells. a-c Cell fractionation and Western blot analyses were performed to assess the nuclear translocation of NF-κB P65. Extracted nucleocytoplasmic proteins were examined using histone H3 as a nuclear protein reference and GAPDH as a cytoplasmic protein reference and analysed with antibodies specific for p65. d Immunofluorescence microscopy analysis of the nuclear translocation of p65 in GES-1 cells. Cells were incubated with p65 antibody and fluorescent secondary antibodies, and nuclei were stained with DAPI. The images were obtained with a fluorescence inverted microscope. The red (representative of the area that contains p65) and blue (representative of the nucleus area that is conjugated to DAPI) images were overlaid to create a merged fluorescence in areas of colocalization (200 x magnification). a GES-1 cells were infected with vacA + Hp strain and Δ vacA Hp strain for 24 h, MOI = 20. The relative protein expression level of p65 (p65/Histone) in nucleus: vacA + Hp strain-infected group vs. Δ vacA Hp strain-infected group, p = 0.046. b and d GES-1 cells were incubated with VacA recombinant protein (65 µg/mL) for 12 h, and GES-1 cells incubated with isovolumetric protein buffer and cell culture medium served as a buffer control group and an untreated normal group, respectively. b The relative protein expression level of p65 (p65/Histone) in nucleus: recombinant VacA protein-incubated group vs. buffer-incubated group, p = 0.039. c GES-1 cells were transfected with pDsRED2-N1-HA/VacA for 48 or 72 h. Note: Comparisons were made between two groups using standard two-tailed Student’s t-test. The figure presents the average of three independent experiments ( n = 3). Data are presented as mean ± SEM. Error bars represent standard error of mean. *: p < 0.05

Article Snippet: Subsequently, the mice were challenged using orogastric infusions of 300 μl of sterile Brucella broth, 5 × 10 9 CFU/ml vacA + Hp strain (the wild-type strain Hp ATCC 26695) or 5 × 10 9 CFU/ml ΔvacA Hp strain ( vacA -KO mutant Hp ATCC 26695) once every 2 days for a total of 9 infusions.

Techniques: Cell Fractionation, Western Blot, Translocation Assay, Immunofluorescence, Microscopy, Incubation, Staining, Fluorescence, Inverted Microscopy, Infection, Expressing, Recombinant, Cell Culture, Control, Transfection, Two Tailed Test

Journal: Molecular Medicine

Article Title: Helicobacter pylori VacA modulates TRAF1-mediated 4-1BB/NF-kappaB axis to induce host apoptosis and chronic inflammatory damage

doi: 10.1186/s10020-025-01349-5

Figure Lengend Snippet: VacA induces apoptosis and inhibits proliferation of GES-1 cells. a The morphological changes in GES-1 cells after infection with the vacA + Hp strain or the Δ vacA Hp strain (MOI = 20) for 24 h were observed by inverted microscopy. Cellular morphological changes were visualized 2 h, 12 h and 24 h after infection by an inverted microscope (magnification × 100). Arrows indicate Helicobacter pylori. b, c GES-1 cells were incubated with VacA recombinant protein at a low concentration (5 µg/ml, 10 µg/ml) for 48 h and at a high concentration (65 µg/mL) for 12 h. In addition, GES-1 cells incubated with isovolumetric protein buffer and cell culture medium served as a buffer control group and an untreated normal group, respectively. Cellular apoptosis was analysed by Annexin V-FITC staining and flow cytometry. (c) Percentage of apoptotic cells were compared between recombinant VacA protein-incubated groups (at concentrations of 5, 10, and 65 µg/mL) and the correspondind buffer-incubated groups, with statistically significant differences ( p = 0.008, p < 0.001, p < 0.001, respectively). d-f GES-1 cells were incubated with VacA recombinant protein (5, 10, and 65 µg/mL) for various durations (12–72 h). Cell viability was assessed by CCK8 assay. The OD450nm levels were compared between recombinant VacA protein-incubated groups (at a concentration of 5 µg/mL) and the corresponding buffer-incubated groups, with statistically significant differences observed at 72 h ( p = 0.045). Levels of OD450nm were compared between recombinant VacA protein-incubated groups (at a concentration of 10 µg/mL) and corresponding buffer-incubated groups at 12, 24, 48, and 72 h, statistically significant differences were observed ( p = 0.008, p = 0.047, p = 0.008, and p = 0.045, respectively). Levels of OD450nm were compared between recombinant VacA protein-incubated groups (at a concentration of 65 µg/mL) and corresponding buffer-incubated groups at 12, 24, and 48 h, statistically significant differences were observed ( p = 0.039, p = 0.005, and p < 0.001, respectively). g-h Cellular apoptosis was analysed by Annexin V-FITC staining and flow cytometry. GES-1 cells were transfected with pDsRED2-N1-HA-VacA in parallel with pDsRED2-N1-HA for 48 h. (h) Percentage of apoptotic cells: pDsRED2-N1-HA/VacA-transfected group vs. empty vector pDsRED2-N1-HA-transfected group, p = 0.047. i Cell proliferation was assessed by an MTT assay. GES-1 cells were transfected with pDsRED2-N1-HA-VacA in parallel with pDsRED2-N1-HA at the indicated time points. Levels of OD490nm were compared between pDsRED2-N1-HA/VacA-transfected groups and empty vector pDsRED2-N1-HA-transfected groups at 24, 48, 72 and 96 h, statistically significant differences were observed ( p = 0.044, p = 0.043, p = 0.038, p < 0.001, respectively). FITC, fluorescein isothiocyanate; PI, propidium iodide. Note: Comparisons were made between two groups using independent sample t tests. The figure presents the average of three independent experiments ( n = 3). Data are presented as mean ± SEM. Error bars represent standard error of mean. *: p < 0.05. **: p < 0.01. ***: p < 0.001

Article Snippet: Subsequently, the mice were challenged using orogastric infusions of 300 μl of sterile Brucella broth, 5 × 10 9 CFU/ml vacA + Hp strain (the wild-type strain Hp ATCC 26695) or 5 × 10 9 CFU/ml ΔvacA Hp strain ( vacA -KO mutant Hp ATCC 26695) once every 2 days for a total of 9 infusions.

Techniques: Infection, Inverted Microscopy, Incubation, Recombinant, Concentration Assay, Cell Culture, Control, Staining, Flow Cytometry, CCK-8 Assay, Transfection, Plasmid Preparation, MTT Assay

Journal: Molecular Medicine

Article Title: Helicobacter pylori VacA modulates TRAF1-mediated 4-1BB/NF-kappaB axis to induce host apoptosis and chronic inflammatory damage

doi: 10.1186/s10020-025-01349-5

Figure Lengend Snippet: TRAF1 promotes the expression of 4-1BB, NF - κ B pathway proteins, apoptosis-related molecules and downstream inflammatory factors in gastric epithelial cells induced by VacA. a, b Western Blot was used to detect the protein expression levels of the target proteins (pIKKα/β, p-P65, p65, TRAF1, 4-1BB, caspase9, Bcl-xl and Bax) after co-culturing TRAF1 stably overexpressing/silenced cell lines with different concentrations of the vacA + Hp strain for 24 h. c, d Western Blot was used to detect the target proteins after co-incubation different concentrations of recombinant VacA protein with TRAF1 stably overexpressing/silenced cell lines for 48 h. e VacA protein (10ug/ml) was incubated with TRAF1 stably overexpressing/silenced gastric epithelial cells for 48 h, using an equal volume of acetic acid buffer as a control. Western blot was used to analyze the expression of various target molecules. f The transfected GES-1 cells were incubated with recombinant VacA protein (10 µg/ml) for 48 h. In addition, the transfected GES-1 cells incubated with isovolumetric protein buffer served as a buffer control group. Western blot was used to detect the effect of TRAF1 silencing or TRAF1 gene overexpression on the expression of the target proteins induced by recombinant VacA protein. g ELISA was used to analyze the secretion of IL-8, IL6, and TNF-αin the TRAF1 stably overexpressing/silenced cells supernatant after co-culturing with vacA + Hp strain or ΔvacA Hp strain at different concentrations for 24 h. Comparison of IL-8, IL-6, and TNF-αlevels between vacA + Hp strain-infected groups and ΔvacA Hp strain-infected groups in HGC27 and MKN74 cells under varying MOI conditions (MOI = 10,20,50,100,200) demonstrated statistically significant differences ( p < 0.001 for all cytokine comparisons across both cell lines and all MOI gradients. * p < 0.05, ** p < 0.01, *** p < 0.001.). When infected with vacA + Hp strain at different multiplicities of infection (MOIs: 10, 20, 50, 100, 200), comparisons of IL-8, IL-6, and TNF-αlevels between TRAF1-OE-HGC27 cells and Vector-HGC27 cells or between shTRAF1-MKN74 cells and shCtrl-MKN74 cells revealed statistically significant differences (all p < 0.05. # p < 0.05, ## p < 0.01, ### p < 0.001.). GAPDH was used as a protein internal reference. Vector-HGC27, TRAF1 OE-HGC27: HGC27 cells infected with control lentivirus or overexpressing lentivirus. shCtrl-MKN74, shTRAF1-MKN74: MKN74 cells infected with control lentivirus or interfering lentivirus. Note: Comparisons were made between two groups using standard two-tailed Student’s t-test. The figure presents the average of three independent experiments ( n = 3). Data are presented as mean ± SEM. Error bars represent standard error of mean. shCtrl, short hairpin control; shTRAF1, short hairpin TRAF1; OE, overexpression

Article Snippet: Subsequently, the mice were challenged using orogastric infusions of 300 μl of sterile Brucella broth, 5 × 10 9 CFU/ml vacA + Hp strain (the wild-type strain Hp ATCC 26695) or 5 × 10 9 CFU/ml ΔvacA Hp strain ( vacA -KO mutant Hp ATCC 26695) once every 2 days for a total of 9 infusions.

Techniques: Expressing, Western Blot, Stable Transfection, Incubation, Recombinant, Control, Transfection, Over Expression, Enzyme-linked Immunosorbent Assay, Comparison, Infection, Plasmid Preparation, Two Tailed Test

Journal: Molecular Medicine

Article Title: Helicobacter pylori VacA modulates TRAF1-mediated 4-1BB/NF-kappaB axis to induce host apoptosis and chronic inflammatory damage

doi: 10.1186/s10020-025-01349-5

Figure Lengend Snippet: TRAF1 promotes the activation of the NF-κB pathway and cell apoptosis in gastric epithelial cells induced by VacA. a VacA recombinant protein (10 µg/mL) was incubated with TRAF1 stably overexpressing/silenced gastric epithelial cells for 48 h, and immunofluorescence was used to analyze NF-κB P65 nuclear translocation. DAPI (blue fluorescence) indicates the cell nucleus, GFP (green fluorescence) indicates the lentiviral fluorescent vector of the stable cell line, and P65 (red fluorescence). The image is magnified 200 times. b Different concentrations of VacA recombinant protein (20, 40 µg/mL) were incubated with TRAF1 stably overexpressing/silenced gastric epithelial cells for 48 h. Cells were stained with Annexin V/PI and apoptosis rates were detected by flow cytometry. Vector-HGC27, TRAF1 OE-HGC27: HGC27 cells infected with control lentivirus or overexpressing lentivirus. shCtrl-MKN74, shTRAF1-MKN74: MKN74 cells infected with control lentivirus or interfering lentivirus.shCtrl, short hairpin control; shTRAF1, short hairpin TRAF1; OE, overexpression

Article Snippet: Subsequently, the mice were challenged using orogastric infusions of 300 μl of sterile Brucella broth, 5 × 10 9 CFU/ml vacA + Hp strain (the wild-type strain Hp ATCC 26695) or 5 × 10 9 CFU/ml ΔvacA Hp strain ( vacA -KO mutant Hp ATCC 26695) once every 2 days for a total of 9 infusions.

Techniques: Activation Assay, Recombinant, Incubation, Stable Transfection, Immunofluorescence, Translocation Assay, Fluorescence, Plasmid Preparation, Staining, Flow Cytometry, Infection, Control, Over Expression

Journal: Molecular Medicine

Article Title: Helicobacter pylori VacA modulates TRAF1-mediated 4-1BB/NF-kappaB axis to induce host apoptosis and chronic inflammatory damage

doi: 10.1186/s10020-025-01349-5

Figure Lengend Snippet: Suppression of the NF-κB signalling pathway counteracts changes in the expression of TRAF1/4-1BB/IL-8 and apoptosis-related proteins induced by VacA in GES-1 cells, resulting in a lower apoptosis rate and increased proliferation. GES-1 cells pretreated with BAY11-7082 (5 µM) were then infected with the vacA + Hp strain at different MOIs (0, 10, 20, 50, 100, and 200) for 24 h or incubated with VacA recombinant protein at a low concentration (5, 10 µg/ml for 48 h) or at a high concentration (65 µg/ml for 12 h). Compared with the VacA recombinant protein incubation group, GES-1 cells incubated with isovolumetric protein buffer and cell culture medium served as a buffer control group and an untreated normal group, respectively. a-d The protein levels of the target molecules (TRAF1, 4-1BB, p65, phospho-p65, Bcl-xl and Bax) were determined by Western blot analysis. GAPDH was used as a loading control. e The secretion level of IL-8 in GES-1 cells after incubation with recombinant VacA protein was measured by ELISA. Comparison of IL-8 levels between recombinant VacA protein-incubated groups (at concentrations of 5, 10, and 65 µg/mL) and recombinant VacA protein + BAY11-7082-incubated groups revealed statistically significant differences at all three concentrations ( p = 0.0045, p < 0.001, and p = 0.0034, respectively). f CCK-8 assay was used to assess the viability of GES-1 cells pretreated with BAY11-7082 and treated with recombinant VacA protein. OD450 levels were compared between groups incubated with 65 µg/mL recombinant VacA protein alone and those incubated with 65 µg/mL recombinant VacA protein + BAY11-7082 at 12, 24, and 48 h, statistically significant differences were observed at all time points ( p = 0.041, p = 0.009, and p = 0.003, respectively). g-h Annexin V-FITC staining coupled with flow cytometry analysis of the apoptosis of GES-1 cells pretreated with BAY11-7082 and treated with VacA recombinant protein. (h) Percentage of apoptotic cells were compared between groups incubated with 65 µg/mL recombinant VacA protein alone and those incubated with 65 µg/mL recombinant VacA protein + BAY11-7082, with statistically significant differences ( p = 0.037). FITC, fluorescein isothiocyanate; PI, propidium iodide. Note: Comparisons were made between two groups using standard two-tailed Student’s t-test. The figure presents the average of three independent experiments ( n = 3). Data are presented as mean ± SEM. Error bars represent standard error of mean. * p < 0.05, ** p < 0.01, *** p < 0.001. BAY, BAY11-7082

Article Snippet: Subsequently, the mice were challenged using orogastric infusions of 300 μl of sterile Brucella broth, 5 × 10 9 CFU/ml vacA + Hp strain (the wild-type strain Hp ATCC 26695) or 5 × 10 9 CFU/ml ΔvacA Hp strain ( vacA -KO mutant Hp ATCC 26695) once every 2 days for a total of 9 infusions.

Techniques: Expressing, Infection, Incubation, Recombinant, Concentration Assay, Cell Culture, Control, Western Blot, Enzyme-linked Immunosorbent Assay, Comparison, CCK-8 Assay, Staining, Flow Cytometry, Two Tailed Test

Journal: Molecular Medicine

Article Title: Helicobacter pylori VacA modulates TRAF1-mediated 4-1BB/NF-kappaB axis to induce host apoptosis and chronic inflammatory damage

doi: 10.1186/s10020-025-01349-5

Figure Lengend Snippet: Effects of BAY11-7082 or 4-1BB blocking/agonist antibodies on TRAF1-mediated 4-1BB, NF-κB pathway activation, and apoptosis-related protein expression in the context of VacA action. a, b HGC27 cells (infected with TRAF1-overexpressing lentivirus or control lentivirus) pretreated with BAY11-7082 (5 µM) were incubated with VacA recombinant protein at different concentrations (5, 10 µg/ml) for 48 h. a Western blot analysis of target molecules (TRAF1, 4-1BB, pIKKα/β, p-P65, and apoptosis-related molecules Caspase9, Bcl-xl, Bax). b ELISA analysis of the secretion of inflammatory factors IL-8, IL6, and TNF-α in the cell supernatant. In Vector-HGC27 cells and TRAF1 OE-HGC27 cells, the levels of IL-8 were compared between groups treated with recombinant VacA protein (5 µg/mL and 10 µg/mL) and groups co-incubated with recombinant VacA protein and BAY11-7082; statistically significant differences were observed at both concentrations in Vector-HGC27 cells ( p = 0.046 and p = 0.039, respectively) and in TRAF1 OE-HGC27 cells ( p = 0.035 and p = 0.024, respectively).In Vector-HGC27 cells, the levels of IL-6 and TNF-α were compared between groups treated with recombinant VacA protein (5 µg/mL and 10 µg/mLL) and groups co-incubated with recombinant VacA protein and BAY11-7082; statistically significant differences were observed for both cytokines at the 10 µg/mL VacA protein concentration (IL-6: p = 0.040; TNF-α: p = 0.031). In TRAF1 OE-HGC27 cells, the levels of IL-6 and TNF-α were compared between groups treated with recombinant VacA protein (5 µg/mL and 10 µg/mL) and groups co-incubated with recombinant VacA protein and BAY11-7082; statistically significant differences were observed for both cytokines at both concentrations (IL-6: p = 0.008 and p = 0.041; TNF-α: p = 0.047 and p = 0.033). c HGC27 cells (infected with overexpressing lentivirus or control lentivirus) pretreated with 4-1BB blocking antibody at different concentrations (5, 10, 20ug/ml) were incubated with VacA recombinant protein (10 µg/ml) for 48 h, Western blot was used to analyze the expression of target molecules in the cells. d MKN74 cells (infected with TRAF1-silencing lentivirus or control lentivirus) pretreated with 4-1BB agonistic antibody at different concentrations (5, 10ug/ml) were incubated with VacA recombinant protein (10 µg/ml) for 48 h, Western blot was used to analyze the expression of target molecules in the cells. Note: Comparisons were made between two groups using standard two-tailed Student’s t-test. The figure presents the average of three independent experiments ( n = 3). Data are presented as mean ± SEM. Error bars represent standard error of mean. * p < 0.05, ** p < 0.01. shCtrl, short hairpin control; shTRAF1, short hairpin TRAF1; OE, overexpression; BAY, BAY11-7082

Article Snippet: Subsequently, the mice were challenged using orogastric infusions of 300 μl of sterile Brucella broth, 5 × 10 9 CFU/ml vacA + Hp strain (the wild-type strain Hp ATCC 26695) or 5 × 10 9 CFU/ml ΔvacA Hp strain ( vacA -KO mutant Hp ATCC 26695) once every 2 days for a total of 9 infusions.

Techniques: Blocking Assay, Activation Assay, Expressing, Infection, Control, Incubation, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Protein Concentration, Two Tailed Test, Over Expression

Journal: Molecular Medicine

Article Title: Helicobacter pylori VacA modulates TRAF1-mediated 4-1BB/NF-kappaB axis to induce host apoptosis and chronic inflammatory damage

doi: 10.1186/s10020-025-01349-5

Figure Lengend Snippet: Constructing models of C57BL/6 mice infected with Hp. a Representative images of Hp, TRAF1 and 4-1BB immunohistochemical staining in gastric mucosal tissue from C57BL/6 mice infected with the vacA + Hp or ΔvacA Hp strains for one month. The black arrows indicate colonized Hp (80×). b Representative immunofluorescence images of the localization of Hp, TRAF1, 4-1BB, and P65 in gastric mucosal tissue of C57BL/6 mice infected with vacA + Hp strains for one month. DAPI (nuclei): blue; Hp: green; TRAF1: red; 4-1BB: orange; P65: pink. Magnification: 40×. c Gastric specimens and HE staining of gastric mucosal tissue from C57BL/6 mice infected with the vacA + Hp and ΔvacA Hp strains for twelve months. Magnification: 30×

Article Snippet: Subsequently, the mice were challenged using orogastric infusions of 300 μl of sterile Brucella broth, 5 × 10 9 CFU/ml vacA + Hp strain (the wild-type strain Hp ATCC 26695) or 5 × 10 9 CFU/ml ΔvacA Hp strain ( vacA -KO mutant Hp ATCC 26695) once every 2 days for a total of 9 infusions.

Techniques: Infection, Immunohistochemical staining, Staining, Immunofluorescence

Journal: Molecular Medicine

Article Title: Helicobacter pylori VacA modulates TRAF1-mediated 4-1BB/NF-kappaB axis to induce host apoptosis and chronic inflammatory damage

doi: 10.1186/s10020-025-01349-5

Figure Lengend Snippet: Expression of TRAF1 and IL-8 of C57BL/6 mice before and after Hp eradication or BAY11-7082 treatment. a C57BL/6 mice were treated with triple anti-Hp therapy six months after infection with the vacA + Hp or ΔvacA Hp strain. Immunohistochemical analysis of TRAF1 protein expression in gastric mucosal tissue from mice in the untreated, 1-month infected, 3-month infected, 6-month infected, 1-month post treatment, and 3-month post treatment groups. The figure shows representative images of immunohistochemical staining for TRAF1 in gastric mucosal tissue. Low magnification: 10×; high magnification: 40×. b, d Effect of BAY11-7082 treatment on TRAF1 expression in mouse gastric mucosal tissue and the serum IL-8 concentration from the mice gavaged with recombinant VacA protein. b, c Immunohistochemical analysis of TRAF1 expression in mouse gastric mucosal tissue from the BSA protein gavage group, protein hydrolysis solution (acetic acid) gavage group, recombinant VacA protein gavage group, and recombinant VacA protein gavage + BAY11-7082 intraperitoneal injection group; c ELISA was performed to quantify the serum IL-8 concentration in each group of mice. Low magnification: 10×; high magnification: 40×. **: P < 0.01. c The integrated optical density (IOD) values of TRAF1: recombinant VacA protein gavage group vs. BSA protein gavage group, p = 0.042; recombinant VacA protein gavage group vs. acetic acid-based buffer control group, p = 0.033; recombinant VacA protein gavage + BAY11-7082 intraperitoneal injection group vs. recombinant VacA protein gavage group, p = 0.046. d The levels of IL-8 of mouse: recombinant VacA protein gavage group vs. BSA protein gavage group, p = 0.039; recombinant VacA protein gavage group vs. acetic acid-based buffer control group, p = 0.035; recombinant VacA protein gavage + BAY11-7082 intraperitoneal injection group vs. recombinant VacA protein gavage group, p = 0.047. Note: Comparisons were made between two groups using standard two-tailed Student’s t-test. The figure presents the average of three independent experiments ( n = 3). Data are presented as mean ± SEM. Error bars represent standard error of mean. * p < 0.05. BAY, BAY11-7082

Article Snippet: Subsequently, the mice were challenged using orogastric infusions of 300 μl of sterile Brucella broth, 5 × 10 9 CFU/ml vacA + Hp strain (the wild-type strain Hp ATCC 26695) or 5 × 10 9 CFU/ml ΔvacA Hp strain ( vacA -KO mutant Hp ATCC 26695) once every 2 days for a total of 9 infusions.

Techniques: Expressing, Infection, Immunohistochemical staining, Staining, Concentration Assay, Recombinant, Injection, Enzyme-linked Immunosorbent Assay, Control, Two Tailed Test