Journal: International Journal of Molecular Sciences

Article Title: Hedyotis diffusa Suppresses Colitis-Associated Colorectal Cancer via Inhibition of the IL-17A-IL-17RA Axis and NF-κB Signaling

doi: 10.3390/ijms27041745

Figure Lengend Snippet: Molecular docking of FA with the IL-17A/IL-17RA complex. The overall docking model shows FA (red) binding at the interface between IL-17A (blue) and IL-17RA (green). The enlarged panel shows a close-up view of the interaction between FA and IL-17A chain B, where hydrogen bonds are indicated by yellow dashed lines.

Article Snippet: Mouse monoclonal antibody against p65 (MAB3026), rabbit polyclonal antibody against IL-17A (bs-1183R), and rabbit monoclonal antibody against CD11b (ab133357) were obtained from Millipore (Temecula, CA, USA), Bioss Antibodies (Woburn, MA, USA), and abcam (Cambridge, UK), respectively.

Techniques: Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Hedyotis diffusa Suppresses Colitis-Associated Colorectal Cancer via Inhibition of the IL-17A-IL-17RA Axis and NF-κB Signaling

doi: 10.3390/ijms27041745

Figure Lengend Snippet: Effects of HD and FA on NF-κB activation, IL-17A production, and granulocyte infiltration in rectal tissues of AOM/DSS-induced CRC mice. Mice were treated with HD (200 mg/kg) or FA (100 mg/kg) for 91 days. Rectal sections were stained with antibody against p65, IL-17A, or CD11b (200× magnification). Scale bar = 100 µm. Representative images are shown. Quantitative analysis of the stained area or cells (%) is displayed below the image. Data are expressed as mean ± SD (n = 8 mice per group). ### p < 0.001 vs. mock group. ** p < 0.01, *** p < 0.001 vs. CRC group.

Article Snippet: Mouse monoclonal antibody against p65 (MAB3026), rabbit polyclonal antibody against IL-17A (bs-1183R), and rabbit monoclonal antibody against CD11b (ab133357) were obtained from Millipore (Temecula, CA, USA), Bioss Antibodies (Woburn, MA, USA), and abcam (Cambridge, UK), respectively.

Techniques: Activation Assay, Staining

Journal: Virulence

Article Title: Gut microbiota-driven IL-17A production by hepatic γδ T cells enhances neutrophil defense against systemic Staphylococcus aureus infection

doi: 10.1080/21505594.2026.2629132

Figure Lengend Snippet: Gut microbiota drive neutrophil recruitment via a IL-17A dependent pathway. The experimental aimed to explore the role of IL-17A in the gut microbiota-mediated enhancement of host resistance against systemic S. aureus infection. The experimental utilized WT mice assigned to No ABX, ABX, and FMT groups, as well as Il17a −/− mice assigned to No ABX and ABX groups. (A) WT mice were i.v . injected with S. aureus USA300, the level of IL-17A was determined in liver tissues by ELISA at 0, 6, 12, and 24 hours post-infection ( n = 5). (B-E) Hepatic IL-17A, KC, MIP-2, and MPO levels in No ABX, ABX, and FMT mice were measured by ELISA at 12 hours post-infection ( n = 5). (F) Schematic shows that Il17a −/− mice were pretreated with either No ABX or ABX containing water. Subsequently, along with the WT-No ABX group, all mice were challenged i.v . with S. aureus USA300. (G) Survival curves of WT-No ABX, Il17a −/− -No ABX, and Il17a −/− - ABX mice after i.v . challenged with S. aureus ( n = 10). (H) Bacterial burden in livers of WT-No ABX, Il17a −/− -No ABX, and Il17a −/− - ABX mice at 12 hours post-infection ( n = 7). (I) Representative contour plots of neutrophils (CD3 − CD11b + Ly6G + ) in liver at 12 hours post-infection. (J) Scatter plots shown absolute number of neutrophils in liver at 12 hours post-infection. ( n = 5). Data are presented as means ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant; one-way ANOVA with Tukey’s for multiple comparisons.

Article Snippet: A single dose of recombinant IL-17A (rIL-17A; MCE), at a dose of 1 μg per mouse based on previous studies [ , ], was dissolved in sterile PBS. rIL-17A or vehicle were mixed with the inoculum of S. aureus and administered intradermally to Tcrδ −/− mice.

Techniques: Infection, Injection, Enzyme-linked Immunosorbent Assay

Journal: Virulence

Article Title: Gut microbiota-driven IL-17A production by hepatic γδ T cells enhances neutrophil defense against systemic Staphylococcus aureus infection

doi: 10.1080/21505594.2026.2629132

Figure Lengend Snippet: Gut microbiota induces IL-17A production by hepatic γδ T cells to promote host systemic defense. The experimental aimed to identify the cellular sources of the key immune factor IL-17A in the liver during gut microbiota-mediated host defense against systemic S. aureus infection. The experimental utilized WT mice assigned to No ABX, ABX, and FMT groups, as well as Tcrδ −/− mice assigned to No ABX, ABX, and ABX+rIL-17A groups. (A) No ABX, ABX, and FMT mice were i.v . challenged with S. aureus USA300, and hepatic Th17 and γδT17 cell levels were measured at 12 hours post-infection. Representative contour plots of hepatic Th17 and γδT17 cells. (B) Scatter plots shown frequencies of hepatic Th17 and γδT17 cells ( n = 5). (C) Schematic shows that Tcrδ −/− mice were pretreated with either No ABX or ABX containing water, or administration with rIL-17A. Subsequently, along with the WT-No ABX group, all mice were challenged i.v . with S. aureus USA300. (D) The level of IL-17A + CD3 + cells in liver were measured in WT-No ABX, Tcrδ −/− -No ABX, and Tcrδ −/− -ABX mice at 12 hours post-infection ( n = 5). Representative contour plots of hepatic IL-17A + CD3 + cells. (E) Scatter plots shown frequencies of hepatic IL-17A + CD3 + cells. (F) ELSIA analyze level of IL-17A in liver. (G) Survival curves of WT-No ABX, Tcrδ −/− -No ABX, Tcrδ −/− -ABX, and Tcrδ −/− -ABX+rIL-17A mice after i.v . challenged with S. aureus ( n = 10). Statistical significance was determined by the log-rank (mantel-cox) test. (H) Bacterial burden in livers from WT-No ABX, Tcrδ −/− -No ABX, Tcrδ −/− -ABX, and Tcrδ −/− -ABX+rIL-17A mice at 12 hours post-infection ( n = 7). (I) Representative images of liver and kidney by H&E staining. Bar: 50 μm. (J) Representative contour plots of neutrophils in liver at 12 hours post-infection. (K) scatter plots shown absolute number of neutrophils in liver at 12 hours post-infection. Data are presented as means ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant; one-way ANOVA with Tukey’s for multiple comparisons.

Article Snippet: A single dose of recombinant IL-17A (rIL-17A; MCE), at a dose of 1 μg per mouse based on previous studies [ , ], was dissolved in sterile PBS. rIL-17A or vehicle were mixed with the inoculum of S. aureus and administered intradermally to Tcrδ −/− mice.

Techniques: Infection, Staining

Journal: Virulence

Article Title: Gut microbiota-driven IL-17A production by hepatic γδ T cells enhances neutrophil defense against systemic Staphylococcus aureus infection

doi: 10.1080/21505594.2026.2629132

Figure Lengend Snippet: Gut microbiota induces IL-17A production by hepatic γδ T cells to promote host systemic defense. (A) The experimental aimed to determine the contribution of L. reuteri WX25, a specific commensal strain derived from the gut microbiota of healthy mice, to host protection against systemic S. aureus infection. The experimental design included the No ABX, Van+Amp, Van+Amp+FMT, and Van+Amp+ L. reuteri WX25 groups. Subsequently, all mice were challenged i.v . with S. aureus USA300. (B) Survival curves of No ABX, Van+Amp, FMT, L. reuteri WX25 mice after i.v . challenged with S. aureus ( n = 10). Statistical significance was determined by the log-rank (mantel-cox) test. (C) bacterial burden in livers from No ABX, Van+Amp, Van+Amp+FMT, and Van+Amp+ L. reuteri WX25 groups at 12 hours post-infection ( n = 6). (D) Representative images of liver and kidney by H&E staining at 12 hours post-infection. Bar: 50 μm. (E) Representative contour plots of γδT17 (CD3 + TCRγ/δ + IL-17A + ) cells in liver at 12 hours post-infection. (F) Scatter plots shown percentage of IL-17A + in γδ T cells in liver at 12 hours post-infection ( n = 5). (G) Representative contour plots of neutrophils in liver at 12 hours post-infection. (H) Scatter plots shown absolute number of neutrophils in liver at 12 hours post-infection ( n = 5). Data are presented as means ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant; one-way ANOVA with Tukey’s for multiple comparisons.

Article Snippet: A single dose of recombinant IL-17A (rIL-17A; MCE), at a dose of 1 μg per mouse based on previous studies [ , ], was dissolved in sterile PBS. rIL-17A or vehicle were mixed with the inoculum of S. aureus and administered intradermally to Tcrδ −/− mice.

Techniques: Derivative Assay, Infection, Staining

Journal: Virulence

Article Title: Gut microbiota-driven IL-17A production by hepatic γδ T cells enhances neutrophil defense against systemic Staphylococcus aureus infection

doi: 10.1080/21505594.2026.2629132

Figure Lengend Snippet: Microbiota-derived indole metabolites promoted host defense against S. aureus systemic infection, involving the γδT17/neutrophil axis. (A) Commensal bacteria isolated from the mouse intestine showed colorimetric evidence of indole metabolites production, as determined by the Kovac’s reagent method. (B) Schematic illustration for the gut microbiota elimination and potential effector metabolites treatment. After antibiotic treatment, mice were treated with indole derivatives (IAld+IAA+IPA) by oral gavage ( i.g .) for 10 days. Subsequently, all mice were challenged i.v . with S. aureus USA300. (C) Survival curves of No ABX, Van+Amp, FMT, L. reuteri WX25 mice after i.v . challenged with S. aureus ( n = 10). Statistical significance was determined by the log-rank (mantel-cox) test. (D) Bacterial burden in livers from No ABX, Van+Amp, FMT, L. reuteri WX25 mice at 12 hours post-infection ( n = 5). (E-H) Hepatic IL-17A, MIP-2, KC, and MPO levels in WT-No ABX, WT-ABX, WT-Indoles, and Tcrδ −/− mice were measured by ELISA at 12 hours post-infection ( n = 5). Data are presented as means ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001; ns, not significant; one-way ANOVA with Tukey’s for multiple comparisons.

Article Snippet: A single dose of recombinant IL-17A (rIL-17A; MCE), at a dose of 1 μg per mouse based on previous studies [ , ], was dissolved in sterile PBS. rIL-17A or vehicle were mixed with the inoculum of S. aureus and administered intradermally to Tcrδ −/− mice.

Techniques: Derivative Assay, Infection, Bacteria, Isolation, Enzyme-linked Immunosorbent Assay

Journal: Immunity & Ageing : I & A

Article Title: Kaempferol alleviates T-cell immunosenescence and inflammaging in aged mice via the SIRT3-LKB1-AMPK-mitophagy pathway

doi: 10.1186/s12979-026-00560-0

Figure Lengend Snippet: SIRT3 is indispensable for kaempferol-induced activation of mitophagy and suppression of inflammation. A Schematic of the dual-lentiviral CRISPR/Cas9 system for SIRT3 knockout in senescent T cells. B Validation of SIRT3 knockout efficiency by immunoblotting in wild-type (WT) cells, WT cells transduced with non-targeting control gRNA (nc gRNA), and WT cells transduced with SIRT3-targeting gRNA (SIRT3 gRNA). (C) Densitometric quantification of SIRT3 protein ( n = 3). D - O Sorted EGFP +/mCherry + T cells (indicating SIRT3-knockout or control) were stimulated with αCD3/αCD28 and treated as indicated. D - G Cells were treated with vehicle or 50 μM Ka for 48 h. D , E Immunoblot analysis (D) and quantification (E) of acetylated LKB1 ( n = 3). F , G Immunoblot analysis (F) and quantification (G) of phosphorylated AMPK (p-AMPK) ( n = 3). H - M Cells were treated with vehicle or 50 μM Ka for 24 h. H , I Immunoblot analysis (H) and quantification (I) of the LC3-II/LC3-I ratio ( n = 3). J , K Immunoblot analysis (J) and quantification (K) of p62 ( n = 3). L , M Immunoblot analysis (L) and quantification (M) of Tom20 ( n = 3). N , O ELISA of (N) TNF-α and (O) IL-17A in culture supernatants ( n = 6). CD3 + T cells were isolated from male mice. For in vitro assays, n denotes independent biological replicates. Data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA with Tukey's post hoc test. * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant

Article Snippet: For SIRT3-knockout experiments, viable EGFP +/mCherry + double-positive T cells were sorted, cultured under the same stimulation/treatment conditions, and supernatants were analysed for IL-17A and TNF-α using ELISA kits (EK217 and EK282EG, Multi Sciences).

Techniques: Activation Assay, CRISPR, Knock-Out, Biomarker Discovery, Western Blot, Transduction, Control, Enzyme-linked Immunosorbent Assay, Isolation, In Vitro