Journal: Advanced Science

Article Title: Western Diet Inhibits FUT7‐Mediated Treg Intestinal Homing to Disrupt the Homeostasis of Intestinal Epithelial Cells in Crohn's Disease

doi: 10.1002/advs.202509541

Figure Lengend Snippet: Upregulation of Fut7 in Tregs improves WD‐induced intestinal barrier disruption in mice. A – L ) Littermate WT mice were divided into four groups ( n = 8/group). They were fed with a normal diet (RD) in group 1 and a western diet (WD) in groups 2–4 for 17 weeks. In week 17, the positive nanoparticle (PN) CD4‐LDP‐Fut7 and the negative nanoparticle (NN) CD4‐LDP‐NC were given by tail intravenous injection at a dose of 240 ng/g once every two days for a total of three times in groups 3 and 4, respectively. ( A ) Fut7 mRNA expression in FACS‐sorted Tregs from the spleen. ( B ) Representative flow cytometric plots of colonic CD25 + Foxp3 + Tregs. ( C ) The frequency of CD25 + Foxp3 + Tregs in CD4 + T cells. ( D ) Colon length. ( E ) Representative gross morphology of the colon. ( F ) After 4 h of fasting, the mice were given FITC‐conjugated dextran by gavage administration for 4 h, and subsequently, the concentration of FITC‐dextran in blood serum was measured. ( G ) Representative intestinal H&E staining. ( H ) Mucosal injury index. ( I ) Inflammatory cell infiltration index. ( J ) Representative immunofluorescence images of Occludin and ZO‐1 immunostaining in colonic tissues. ( K, L ) MFI of Occludin and ZO‐1 was quantified by ImageJ software. The data shown are representative of three independent experiments. ( A, C, D, F, K, L ) one‐way ANOVA or ( H, I ) Kruskal‐Wallis test; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant. FACS, fluorescence‐activated cell sorting; MFI, mean fluorescence intensity; WT, wild‐type.

Article Snippet: A naive CD4 + T cell adoptive transfer model was established using Rag2 −/− mice purchased from Cyagen Biosciences (Guangzhou, China) according to a published procedure.

Techniques: Disruption, Western Blot, Injection, Expressing, Concentration Assay, Staining, Immunofluorescence, Immunostaining, Software, Fluorescence, FACS

Journal: Advanced Science

Article Title: Western Diet Inhibits FUT7‐Mediated Treg Intestinal Homing to Disrupt the Homeostasis of Intestinal Epithelial Cells in Crohn's Disease

doi: 10.1002/advs.202509541

Figure Lengend Snippet: Specific deletion of Fut7 in Tregs deteriorates TNBS‐induced intestinal barrier disruption in mice. A ) fut7 fl/fl foxp3 cre mice with conditional knockout (CKO) of the fut7 gene in Tregs were generated as shown in the schematic diagram. B ) The knockout efficiency was determined by Western blot analysis of Fut7 expression in FACS‐sorted splenic Tregs. C ) Representative flow cytometric plots of colonic CD25 + Foxp3 + Tregs. D ) The frequency of CD25 + Foxp3 + Tregs in CD4 + T cells. E–K ) CKO and fut7 fl/fl littermates were intrarectally injected with TNBS to induce colitis (n = 6–8/group). ( E ) Mouse weight changes. ( F ) Colon length. ( G ) Representative gross morphology of the colon. ( H ) After 4 h of fasting, the mice were given FITC‐conjugated dextran by gavage administration for 4 h, and subsequently, the concentration of FITC‐dextran in blood serum was measured. ( I ) Representative intestinal H&E staining. ( J ) Mucosal injury index. ( K ) Inflammatory cell infiltration index. Data shown in C–K are representative of three independent experiments. ( D ) unpaired Student's t test, ( E , F, H ) one‐way ANOVA, or ( J, K ) Kruskal‐Wallis test; * P < 0.05, ** P < 0.01, **** P < 0.0001; ns, not significant. FACS, fluorescence‐activated cell sorting; TNBS, 2,4,6‐trinitrobenzenesulfonic acid.

Article Snippet: A naive CD4 + T cell adoptive transfer model was established using Rag2 −/− mice purchased from Cyagen Biosciences (Guangzhou, China) according to a published procedure.

Techniques: Disruption, Knock-Out, Generated, Western Blot, Expressing, Injection, Concentration Assay, Staining, Fluorescence, FACS

Journal: Advanced Science

Article Title: Western Diet Inhibits FUT7‐Mediated Treg Intestinal Homing to Disrupt the Homeostasis of Intestinal Epithelial Cells in Crohn's Disease

doi: 10.1002/advs.202509541

Figure Lengend Snippet: FUT7 expression in Tregs and CD15s + Tregs is decreased in patients with active CD. A ) Fut7 mRNA expression in FACS‐sorted Tregs from the blood of patients with active CD (n = 6), quiescent CD (n = 6), and healthy controls (n = 6). B–E ) PBMCs were isolated from the blood of patients with active CD (n = 8), quiescent CD (n = 8), and healthy controls (n = 8) by density centrifugation, and then CD4 + CD25 + CD127 low Tregs and CD15s + Tregs were analyzed by flow cytometry. ( B, D ) Representative flow cytometric plots. ( C ) The frequency of CD25 + CD127 low Tregs in CD4 + T cells. ( E ) The frequency of CD15s + Tregs in Treg cells. ( F to H ) Fresh colonic mucosa from active CD ( n = 9), quiescent CD ( n = 9), and healthy controls ( n = 9) were fixed in 4% paraformaldehyde and subjected to immunofluorescence staining using anti‐human antibodies targeting Foxp3 and CD15s. ( F ) Representative immunofluorescence images. ( G, H ) The number of indicated Treg subsets. ( A, C, E, G, H ) One‐way ANOVA; ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant. FACS, fluorescence‐activated cell sorting; PBMC, peripheral blood mononuclear cell.

Article Snippet: A naive CD4 + T cell adoptive transfer model was established using Rag2 −/− mice purchased from Cyagen Biosciences (Guangzhou, China) according to a published procedure.

Techniques: Expressing, Isolation, Centrifugation, Flow Cytometry, Immunofluorescence, Staining, Fluorescence, FACS

Journal: Frontiers in Oncology

Article Title: Guanylate cyclase-C Signaling Axis as a theragnostic target in colorectal cancer: a systematic review of literature

doi: 10.3389/fonc.2023.1277265

Figure Lengend Snippet: GUCY2C-associated studies included in the systematic review.

Article Snippet: • Vaccine: Investigating a prime-boost approach involving a chimeric adenoviral vector (Ad5.F35) engineered to overcome pre-existing immunity, followed by recombinant Lm to enhance immune response toward the gastrointestinal cancer antigen GUCY2C. , • Vaccines: Ad-GUCY2C • Adenovirus expressing mouse GUCY2C1-429 fused to the influenza HA107-119 CD4+ T-cell epitope • Lm-GUCY2C and Lm-LacZ • mouse macrophage cell line J774A.1 (ATCC) • BALB/cJ mice , • In vitro infections • Immunizations • Ad5-neutralizing immunity studies • IFNγ ELISpot assay • Intracellular cytokine staining • In vivo tumor studies • Safety studies • Blood chemistry and cytokine analyses • Western blot , • Immunization with both heterologous Ad-GUCY2C and Lm-GUCY2C enhances CD8+ T-cell responses specific to GUCY2C and boosts antitumor immune activity. • Previous exposure to Ad5 restricts the effectiveness of Ad5.F35+Lm immunization protocols, whereas prior Lm exposure does not impose such limitations. • Alterations in the qualitative characteristics of the CD8+ T-cell population after a prime-boost vaccination regimen. • Lm-GUCY2C could potentially be employed to enhance GUCY2C-specific immune responses in patients undergoing clinical trials with adenovirus-based GUCY2C vaccines, aiming to prevent or manage recurrent GI cancer. , ( ) .

Techniques: Amplification, Expressing, Quantitative RT-PCR, Radioactivity, Activity Assay, Modification, Purification, Labeling, In Vivo, Mouse Assay, In Vitro, Clinical Proteomics, Viability Assay, Release Assay, Fluorescence, Staining, Immunofluorescence, Enzyme-linked Immunospot, Luciferase, Cell Culture, Activation Assay, Histopathology, Immunohistochemistry, Drug discovery, Western Blot, Isolation, Sequencing, Transduction, Variant Assay, Generated, Ex Vivo, Conjugation Assay, Flow Cytometry, Cytotoxicity Assay, Injection, Imaging, Mutagenesis, Protein-Protein interactions, Knock-Out, Transformation Assay, Plasmid Preparation, Control, Neutralization, Vaccines, Marker, Cell Isolation, Adoptive Transfer Assay, Transgenic Assay, Binding Assay, Cytotoxic T Lymphocyte Assay, Animal Model, Digital PCR, Gene Expression, Positron Emission Tomography, Comparison, Stability Assay, Immunopeptidomics, Recombinant, RNA Sequencing, Cloning, CRISPR, Chromatin Immunoprecipitation

Journal: eBioMedicine

Article Title: The therapeutic effects of the VEGF decoy receptor fusion protein VEGF-Grab in chronic inflammatory diseases

doi: 10.1016/j.ebiom.2026.106216

Figure Lengend Snippet: Inhibitory effects of PB101 on angiogenesis. ( A ) Endothelial cell chemotactic migration assay. EA.hy926 cells (1 × 10 5 , n = 3) were loaded in the upper chambers of Transwell inserts and chemoattracted by VEGF (100 ng/mL) or PlGF-2 (40 ng/mL) with or without PB101 (2 or 5 μg/mL) in the lower chambers. The cells that migrated to the underside of the Transwell chamber were manually counted, and the results are presented as a percentage of the untreated control (none). ( B ) Wounding migration of endothelial cells. EA.hy926 cells (4 × 10 5 , n = 5) seeded on a 6-well plate were wounded using pipette tips and treated with VEGF (200 ng/mL) or PlGF-2 (100 ng/mL) with or without PB101 (2 or 5 μg/mL). The cells that crossed the reference lines (red dashed lines) were counted as migrated cells. ( C ) Capillary tube formation by endothelial cells. EA.hy926 cells (3 × 10 4 , n = 3–4) were seeded on a Matrigel-coated 96-well plate and incubated with VEGF (500 ng/mL) or PlGF-2 (500 ng/mL) with or without PB101 (10 or 50 μg/mL). The tube formation area was measured and is presented as an arbitrary unit (a.u.). The cellular images are representative of three independent experiments. Scale bar, 200 μm. The bar graphs present the mean and SEM. The p-values were determined via one-way ANOVA with Tukey's multiple comparisons test. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001 vs. untreated controls. # p < 0.05, ## p < 0.01 vs. PlGF- or VEGF-treated positive controls. ( D and E ) Suppression of recombinant PlGF-induced angiogenesis by PB101 in a Matrigel plug assay. C57BL/6 mice (n = 3, total 12 mice) were injected subcutaneously with Matrigel mixed with VEGF (500 ng/mL) or recombinant PlGF-2 (200 ng/mL) with or without PB101 (50 μg/mL) (D). Mice were injected with vehicle or PB101 (50 mg/kg) every day. To assess the effects of PlGF-overexpressing T cells, CD4 + T cells isolated from PlGF transgenic (PlGF Tg) mice were stimulated with anti-CD3/CD28 Abs for 48 h. PlGF Tg CD4 + T cells (5 × 10 5 ) and their culture supernatants were incorporated into Matrigel with or without PB101 (50 μg/mL) (E). C57BL/6 mice (n = 3) were subcutaneously injected with these Matrigel plugs and received daily injections of Fc vehicle or PB101 (50 mg/kg). After 14 days, the vascularity of the Matrigel plugs was evaluated through visual assessment and H&E staining of the tissue sections. Scale bars, 500 nm (top) for Matrigel plugs, and 100 (middle) and 1000 (bottom) μm for H&E. The boxed areas in the upper panels of the H&E-stained images are shown at higher magnification in the lower panels. ( F ) Immunohistochemical staining of the Matrigel plugs was performed using an anti-F4/80 Ab to assess macrophage infiltration. The macrophages were manually counted in three randomly selected fields per section of total nine sites per group. Scale bars, 100 μm. The bar graphs present the mean and SEM. ∗∗∗∗p < 0.0001 vs. untreated controls. ### p < 0.001 and #### p < 0.0001 vs. PB101-untreated positive controls. The p-values were determined via Kruskal–Wallis with Dunn's multiple comparisons test.

Article Snippet: Immune cells were isolated from the spleens and LNs of C57BL/6 mice and subsequently purified by magnetic separation using a naïve CD4 + T cell isolation kit (catalog 130-104-453; Miltenyi Biotech, Bergisch Gladbach, Germany).

Techniques: Migration, Control, Transferring, Incubation, Recombinant, Matrigel Assay, Injection, Isolation, Transgenic Assay, Staining, Immunohistochemical staining

Journal: eBioMedicine

Article Title: The therapeutic effects of the VEGF decoy receptor fusion protein VEGF-Grab in chronic inflammatory diseases

doi: 10.1016/j.ebiom.2026.106216

Figure Lengend Snippet: VEGF-Grab inhibits the differentiation and function of Th17 cells in vitro . ( A ) The inhibitory effect of VEGF-Grab against Th17 differentiation was measured. Th17 cell differentiation (1.5 × 10 5 , n = 3) was induced by adding mIL-6 (20 ng/mL), hTGF-β (1 ng/mL), anti–IFN–γ (1 μg/mL), anti-IL-4 (1 μg/mL), and anti-IL-2 (1 μg/mL) during stimulation with anti-CD3/CD28 antibodies (0.5 μg/mL). Pathogenic Th17 cell differentiation was supplemented with mIL-1β (20 ng/mL) and hIL-23 (30 ng/mL) in Th17 cell condition. Either Fc control (Fc, 50 μg/mL), PB101, or PB102 (25 or 50 μg/mL) was added, and Th17 cells were quantified using flow cytometric analyses. ( B ) The suppressive effects of VEGF-Grab on Th17 cell function (n = 3). Fully differentiated Th17 cells were re-stimulated with Th17 differentiation condition in the presence of Fc control (Fc, 50 μg/mL), PB101, or PB102 (25 or 50 μg/mL). Total Th17 cells and pathogenic Th17 cells were enumerated in flow cytometric analyses. ( C ) The inhibitory effects of VEGF-Grab against the antigen-specific Th17 differentiation of OT-II CD4 + T cells (n = 3). OT-II T cells were stimulated with OVA peptide 323-338 , and Th17 cell or pathogenic Th17 cell differentiation was induced in the conditions given in (A) with either Fc control (Fc, 50 μg/mL), PB101, or PB102 (25 or 50 μg/mL). The percentage of total Th17 cells and pathogenic Th17 cells was evaluated in flow cytometric analyses. Data are presented as the mean and SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001 vs. Fc vehicle. The p-values were determined via the unpaired one-way ANOVA with Tukey's multiple comparisons test (A and C) and paired one-way ANOVA with Tukey's multiple comparisons test (B).

Article Snippet: Immune cells were isolated from the spleens and LNs of C57BL/6 mice and subsequently purified by magnetic separation using a naïve CD4 + T cell isolation kit (catalog 130-104-453; Miltenyi Biotech, Bergisch Gladbach, Germany).

Techniques: In Vitro, Cell Differentiation, Control, Cell Function Assay

Journal: eBioMedicine

Article Title: The therapeutic effects of the VEGF decoy receptor fusion protein VEGF-Grab in chronic inflammatory diseases

doi: 10.1016/j.ebiom.2026.106216

Figure Lengend Snippet: Therapeutic effects of VEGF-Grab in inflammatory and autoimmune arthritis models. ( A ) A schematic representation of AIA induction and the drug treatment schedule. ( B ) C57BL/6 mice (n = 5, total 10 mice) were subcutaneously injected on the right foot with 50 μL of complete Freund's adjuvant (CFA). Beginning on the day of arthritis induction, the mice were injected with vehicle or PB102 (4 mg/kg) daily. The thickness of the ankle and footpad was measured and presented as a percentage of their initial values before the induction of arthritis. Scale bars, 50 mm. The graphs are presented as the mean and SEM. ∗∗∗p < 0.001 vs. vehicle controls. The p-values were determined via two-way ANOVA. ( C ) Representative images of H&E-stained tissue sections. The rectangular areas with a black line in the upper panels are magnified in the lower panels. The levels of inflammation, synovial proliferation, and bone destruction were evaluated by histological analysis of the H&E-stained tissue sections. The bar graphs present the mean and SEM. ∗p < 0.05 vs. vehicle controls. The p-values were determined via the Mann–Whitney U test. ( D ) Detection of IL-17 with ELISA in the culture supernatant of CD4 + T cell isolated from AIA induced mice stimulated with anti-CD3/CD28 antibodies (0.5 or 1 μg/mL) for 3 days. Data are presented as the mean and SEM. ∗p < 0.05 vs. each vehicle-treated group. Statistical significance was determined by Kruskal–Wallis test with Dunn's multiple comparisons. ( E ) A schematic representation of CIA induction and the drug treatment schedule. DBA-1 mice (n = 10, total 30 mice) were immunised with type II bovine collagen. After secondary immunisation, the mice were treated with vehicle or PB102 (4 or 10 mg/kg) twice a week. ( F ) The clinical scores of the CIA mice were visually determined. ∗∗∗p < 0.001 vs. vehicle. ## p < 0.01 vs PB102 (4 mg/kg)-treated group. The p-values were determined via two-way ANOVA. ( G and H ) Analyses of histopathological features (H) and pannus formation (G) in the arthritic limbs of CIA mice. Representative images of H&E-stained tissue sections are presented in the left panel, and the results of the histological analysis are in the right panel. Scale bars, 200 μm. The data are presented as the mean and SEM. The p-values were determined a one-way analysis of variance (ANOVA) with Tukey's multiple comparisons test or Kruskal–Wallis with Dunn's multiple comparisons test was used. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 vs. vehicle. # p < 0.05 vs. PB102 (10 mg/kg). ( I and J ) Immunohistochemical analysis of macrophage infiltration and angiogenesis. Tissue sections from vehicle controls (n = 4) and PB102-treated groups (n = 5) were subjected to immunohistochemical staining for F4/80 for macrophages (I), and CD31 for blood vessels (J). Representative images are shown (left), and the graphs (right) display the quantification of F4/80-positive cells relative to total cells and blood vessel numbers, respectively. Scale bars, 100 μm (I, upper panel), 20 μm (I, lower panel), and 50 μm (J). The data are presented as the mean and SEM. ∗p < 0.05 and ∗∗∗p < 0.001 vs. vehicle controls. ( K ) Detection of IL-17 in the culture supernatant of CD4 + T cell isolated from mice with CIA. CIA mice were administered vehicle (n = 6) or PB102 at 10 mg/kg (n = 4) according to the protocol described in (E). Splenic CD4 + T cells were isolated 5 weeks after CIA induction and stimulated with anti-CD3/CD28 antibodies (0.5 or 1 μg/mL) for 3 days. IL-17 production in the culture supernatants was then measured by ELISA. Data are presented as the mean and SEM. ∗p < 0.05 and ∗∗p < 0.01 vs. each vehicle-treated group. Statistical significance was determined by Kruskal–Wallis test with Dunn's multiple comparisons.

Article Snippet: Immune cells were isolated from the spleens and LNs of C57BL/6 mice and subsequently purified by magnetic separation using a naïve CD4 + T cell isolation kit (catalog 130-104-453; Miltenyi Biotech, Bergisch Gladbach, Germany).

Techniques: Injection, Adjuvant, Staining, MANN-WHITNEY, Enzyme-linked Immunosorbent Assay, Isolation, Immunohistochemical staining

Journal: eBioMedicine

Article Title: The therapeutic effects of the VEGF decoy receptor fusion protein VEGF-Grab in chronic inflammatory diseases

doi: 10.1016/j.ebiom.2026.106216

Figure Lengend Snippet: Therapeutic effects of VEGF-Grab (PB102) in an animal model of MS. ( A ) A schematic representation of EAE induction and the drug treatment schedule. C57BL/6 mice (n = 5, total 15 mice) were subcutaneously immunised with a CFA/MOG emulsion. Beginning on day 8 after the induction, the mice were treated with Fc control (1 mg/kg) or PB101 (1 mg/kg) twice a week. ( B ) Measurements of EAE pathologic scores for 24 days. ( C ) Analyses of CNS-infiltrated immune cells by flow cytometry. The frequencies of IL-17 and GM-CSF, or IL-17 and IFN-γ double-positive CD4 + T cells, known as pathogenic Th17 cells, and neutrophils are shown. ( D ) Analyses of pathogenic Th17 cells in draining LNs. ( E ) Quantification of inflammatory cytokine secretion by CNS-infiltrated immune cells after 3 days of culture with anti-CD3/CD28 Abs (2 μg/mL) stimulation. Data are presented as the mean and SEM. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗∗p < 0.0001 vs. negative control. # p < 0.05, ## p < 0.01, and ### p < 0.001 vs. Fc vehicle. The p-values were determined via one-way ANOVA with Tukey's multiple comparisons test.

Article Snippet: Immune cells were isolated from the spleens and LNs of C57BL/6 mice and subsequently purified by magnetic separation using a naïve CD4 + T cell isolation kit (catalog 130-104-453; Miltenyi Biotech, Bergisch Gladbach, Germany).

Techniques: Animal Model, Emulsion, Control, Flow Cytometry, Negative Control