Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: IFN-γ suppresses tumor growth and invasion. (A) Cytokine profiling of co-culture supernatants via ELISAs: IFN-γ, IL-1β, IL-6, IL-10, TGF-β and TNF-α. (B-D) Spatial expression patterns of IFN-γ. (B) Immunofluorescence imaging of the invasive front in SSIT, showing DAPI (blue), IBA-1 + macrophages (red) and IFN-γ + signals (green), and grayscale intensity distribution. (C) Immunofluorescence imaging of TIM and NIM, showing DAPI (blue), IBA-1 + macrophages (red) and IFN-γ + signals (green). (D) Quantification of relative IFN-γ expression in TIM and NIM. (E) Representative Ki-67 immunohistochemistry images of SSIT cases stratified into IFN-γ-high and IFN-γ-low groups (n=5 each; median split). (F) Quantification of Ki-67 index comparing the two groups. (G) EdU staining demonstrating dose-dependent suppression of TtT/GF pituitary adenoma cell proliferation by IFN-γ (0–100 ng/ml; 48 h). (H) Representative flow cytometry histograms for cell cycle analysis of cells treated with IFN-γ (0–100 ng/ml) in the absence (0 µM) or presence (5 µM) of ruxolitinib. (I) Stacked bar plot showing the percentages of cells in the G 1 , S and G 2 /M phases under the same treatment conditions. (A) One-way ANOVA with Tukey's post hoc multiple comparisons test. (D and F) Unpaired two-tailed Student's t-test. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. CTRL, control; DMC, digested mucosal culture; EdU, 5-ethynyl-2′-deoxyuridine; IBA-1, ionised calcium binding adaptor molecule 1; MTC, mucosal tissue culture; NIM, non-invaded mucosa; ns, not significant; PE-A, phycoerythrin-area; SSIT, sphenoid sinus-invasive tumor; TIM, tumor-invaded mucosa.

Article Snippet: Human IFN-γ, IL-1β, IL-6, IL-10, TGF-β and TNF-α levels were quantified using commercial ELISA kits (Human IFN-γ ELISA kit, cat. no. E-EL-H0108; Human IL-1β ELISA kit, cat. no. E-EL-H0149; Human IL-6 ELISA kit, cat. no. E-EL-H0102; Human IL-10 ELISA kit, cat. no. E-EL-H0103; Human TGF-β ELISA kit, cat. no. E-EL-H0110; Human TNF-α ELISA kit, cat. no. E-EL-H0109; Wuhan Elabscience Biotechnology Co., Ltd.).

Techniques: Co-Culture Assay, Expressing, Immunofluorescence, Imaging, Immunohistochemistry, Staining, Flow Cytometry, Cell Cycle Assay, Two Tailed Test, Control, Binding Assay

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: Elevated IgG levels drive macrophage M2-to-M1 reprogramming. (A) Sphenoid sinus-invasive tumor cases stratified into CD19-high (n=5) and CD19-low (n=5) groups based on the cohort median of CD19 + B cell density, with (B) quantitative analyses of macrophage polarization (M1-like versus M2-like). (C) Dural-invasive tumor and non-invasive tumor cases stratified into IgG-high (n=27) and IgG-low (n=26) groups based on the cohort median of relative IgG immunohistochemistry staining intensity, with (D) quantitative analyses of M1-like/M2-like macrophage proportions. (E and F) RAW264.7 macrophages were pre-polarized with IL-4 (20 ng/ml) or with lipopolysaccharide (100 ng/ml) plus IFN-γ (20 ng/ml) for 24 h, followed by IgG (10 µg/ml) exposure. Relative (E) IL-6 and (F) TNF-α mRNA expression in RAW264.7 macrophages pre-polarized to M0, M1 or M2 states. (G) Representative flow cytometric cell-cycle profiles of TtT/GF cells following the indicated treatments. (H) Stacked bar plot summarizing the percentages of cells from (G) in G 1 , S and G 2 /M phases. (I) Representative images from the scratch wound assay at 0, 24, 48 and 72 h under the indicated treatments. (J) Quantification of scratch wound closure. (K) Representative western blot images showing total STAT1, p-STAT1, total STAT3, p-STAT3 and β-actin levels in cells treated with IFN-γ (100 ng/ml), IL-6 (100 ng/ml), IFN-γ + IL-6 (50 ng/ml each), ruxolitinib (5 µM) or IFN-γ + IL-6 (50 ng/ml each) plus ruxolitinib (5 µM), as indicated. (L) Densitometric semi-quantification of p-STAT1/STAT1 (ratio). (B and D) Unpaired two-tailed Student's t-test. (E, F, J and L) One-way ANOVA with Tukey's post hoc multiple comparisons test. *P<0.05, ***P<0.001, ****P<0.0001. CTRL, control; IBA-1, ionised calcium binding adaptor molecule 1; ns, not significant; p-, phosphorylated; PE-A, phycoerythrin-area.

Article Snippet: Human IFN-γ, IL-1β, IL-6, IL-10, TGF-β and TNF-α levels were quantified using commercial ELISA kits (Human IFN-γ ELISA kit, cat. no. E-EL-H0108; Human IL-1β ELISA kit, cat. no. E-EL-H0149; Human IL-6 ELISA kit, cat. no. E-EL-H0102; Human IL-10 ELISA kit, cat. no. E-EL-H0103; Human TGF-β ELISA kit, cat. no. E-EL-H0110; Human TNF-α ELISA kit, cat. no. E-EL-H0109; Wuhan Elabscience Biotechnology Co., Ltd.).

Techniques: Immunohistochemistry, Staining, Expressing, Scratch Wound Assay Assay, Western Blot, Two Tailed Test, Control, Binding Assay

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: Anti-CD47 mAb enhances ADCP to suppress tumor cell proliferation. (A) Immunofluorescence staining of CD47 (red) and DAPI (blue) in a representative subset of non-invasive tumor, dural-invasive tumor and sphenoid sinus-invasive tumor cases (n=10 per group). (B) Paired comparison of CD47 fluorescence intensity at the IF versus the TC. (C) RAW264.7 macrophages were pre-polarized with IL-4 (20 ng/ml) or with lipopolysaccharide (100 ng/ml) plus IFN-γ (20 ng/ml) for 24 h, followed by anti-CD47 mAb (10 µg/ml) treatment for 12 h. Quantitative PCR was used to analyze polarization/activation markers. (D) Schematic illustrating anti-CD47 mAb-mediated blockade of the CD47-SIRPα axis and enhancement of ADCP. (E) EdU assay of TtT/GF cell proliferation in a Transwell co-culture with anti-CD47 mAb-treated polarized macrophages. (F) Quantification of EdU-positive cells. (G) Representative microscopy images and flow cytometry plots showing macrophage phagocytosis of pHrodo™ Red-labeled GFP-TtT/GF cells. (H) Quantification of phagocytosis (%). (B) Paired two-tailed Student's t-test. (C, F and H) One-way ANOVA with Tukey's post hoc multiple comparisons test. **P<0.01, ***P<0.001, ****P<0.0001. ADCP, antibody-dependent cellular phagocytosis; Arg-1, arginase 1; EdU, 5-ethynyl-2′-deoxyuridine; FcγR, Fcγ receptor; GFP, green fluorescent protein; IF, invasive front; mAb, monoclonal antibody; NOS2, nitric oxide synthase 2; ns, not significant; PE, phycoerythrin; SIRPα, signal regulatory protein-α; SSCA, side scatter area; TC, tumor core.

Article Snippet: Human IFN-γ, IL-1β, IL-6, IL-10, TGF-β and TNF-α levels were quantified using commercial ELISA kits (Human IFN-γ ELISA kit, cat. no. E-EL-H0108; Human IL-1β ELISA kit, cat. no. E-EL-H0149; Human IL-6 ELISA kit, cat. no. E-EL-H0102; Human IL-10 ELISA kit, cat. no. E-EL-H0103; Human TGF-β ELISA kit, cat. no. E-EL-H0110; Human TNF-α ELISA kit, cat. no. E-EL-H0109; Wuhan Elabscience Biotechnology Co., Ltd.).

Techniques: Immunofluorescence, Staining, Comparison, Fluorescence, Real-time Polymerase Chain Reaction, Activation Assay, EdU Assay, Co-Culture Assay, Microscopy, Flow Cytometry, Labeling, Two Tailed Test

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: Summary graphic illustration. This illustration summarizes the proposed model during pituitary adenoma invasion. The tumor invasive front abuts an intact sphenoid sinus mucosa, forming a distinct boundary. The mucosal compartment is enriched for ionised calcium binding adaptor molecule 1-positive macrophages with an M1-like predominance and IgG-high B cells. B cell-derived IgG promotes M2-to-M1 macrophage reprogramming, while coordinated IFN-γ and IL-6 production establishes a tumor-suppressive cytokine gradient that decreases from mucosa toward the tumor core, constraining proliferation and migration via JAK-STAT1 activation. Therapeutically, anti-CD47 monoclonal antibody blocks the CD47-SIRPα ‘don't-eat-me’ axis and augments antibody-dependent cellular phagocytosis, highlighting a strategy for immune checkpoint-targeted therapy that may complement surgical management. FcR, Fc receptor; JAK, Janus kinase; mAb, monoclonal antibody; p-, phosphorylated; SIRPα, signal regulatory protein-α.

Article Snippet: Human IFN-γ, IL-1β, IL-6, IL-10, TGF-β and TNF-α levels were quantified using commercial ELISA kits (Human IFN-γ ELISA kit, cat. no. E-EL-H0108; Human IL-1β ELISA kit, cat. no. E-EL-H0149; Human IL-6 ELISA kit, cat. no. E-EL-H0102; Human IL-10 ELISA kit, cat. no. E-EL-H0103; Human TGF-β ELISA kit, cat. no. E-EL-H0110; Human TNF-α ELISA kit, cat. no. E-EL-H0109; Wuhan Elabscience Biotechnology Co., Ltd.).

Techniques: Binding Assay, Derivative Assay, Migration, Activation Assay

Journal: Redox Biology

Article Title: A novel photosensitizer-based photodynamic therapy reprograms the Kynurenine–AhR axis to boost antitumor immunity in breast cancer

doi: 10.1016/j.redox.2026.104171

Figure Lengend Snippet: DTP-PDT attenuates the IDO–Kyn–AhR axis and relieves immune-suppression in the TME. (A-D) Targeted metabolomics analysis of Trp, Kyn, QA, and 5-HT in cell samples. (E) Targeted metabolomics of Kyn in tumor tissue samples. (F) Levels of Kyn in culture supernatants after the indicated various treatments under IFN-γ priming (n = 3). (G) Representative immunofluorescence images of tumor sections stained for CD3 (green), AhR (red), and DAPI (blue). Scale bar = 20 μm. (H) RT-qPCR analysis of Cyp1a1, Cyp1b1, and Ahrr mRNA expression in tumor-infiltrating CD3 + T cells (n = 4). (I-J) Proportion of intratumoral CD8 + T cells (gated on CD3 + T cells, n = 5). (K-L) Proportion of intratumoral Treg cells (gated on CD3 + CD4 + Foxp3 + T cells, n = 5). (M) Representative immunofluorescence images of tumor sections stained for CD3 (green), AhR (red), and DAPI (blue) in the Kyn rescue experiment. Scale bar = 20 μm. (N) RT-qPCR analysis of Cyp1a1, Cyp1b1, and Ahrr mRNA expression in tumor-infiltrating CD3 + T cells from the Kyn rescue experiment (n = 4). (O–P) Proportion of intratumoral CD8 + T cells in the Kyn rescue experiment (gated on CD3 + T cells, n = 5). (Q-R) Proportion of intratumoral Treg cells in the Kyn rescue experiment (gated on CD3 + CD4 + Foxp3 + T cells, n = 5). Data are shown as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: 4T1 cells (5 × 10 5 cells/well) were seeded in 6-well plates and allowed to adhere for 24 h. To activate IDO1 activity, cells were pretreated with recombinant mouse interferon-γ (IFN-γ) (50 ng/mL, 485-MI, R&D System) for 24 h. After IFN-γ priming, cells were treated with DTP-PDT and/or the IDO1 inhibitor (NLG919, Aladdin).

Techniques: Immunofluorescence, Staining, Quantitative RT-PCR, Expressing

Journal: Bioactive Materials

Article Title: Carrier free oral Co-delivery of atorvastatin via baicalein-copper-network for atherosclerosis therapy through senescence reversal and multi-mechanistic synergy

doi: 10.1016/j.bioactmat.2025.12.036

Figure Lengend Snippet: Reactive oxygen species scavenging capacity of CMA. (A) H 2 O 2 (100 μM) scavenging capacity CMA at 2 h. (B) O 2 − scavenging capacity of CMA at 40 min. (C) •OH scavenging capacity of CMA at 1 min. (D) DPPH (100 μg mL -1 ) scavenging capacity of CMA at 1 h. (E) Fluorescence images of RAW264.7 cells treated with CMA and LPS + IFN-γ for 24 h. Intracellular ROS were stained with DCFH-DA (Green), and nuclei were stained with DAPI (blue). Scale bar = 50 μm. (F) Quantitative analysis of mean DCFH-DA fluorescence intensity. Flow cytometric analysis of ROS expression in: (G) RAW264.7: CMA + LPS; (H) RAW264.7: CMA; (I) SMC: CMA; (J) HUVEC: CMA; (K) RAW264.7: CMA + BAPTA; (L) SMC: CMA + BAPTA; (M) HUVEC: CMA + BAPTA. All treatments were conducted for 24 h. Data are presented as mean ± SD (n = 3–6; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001).

Article Snippet: Baicalein (BAI), copper chloride (CuCl 2 ·2H 2 O), and Atorvastatin (ATV) were purchased from Macklin Inc. Lipopolysaccharides (LPS), recombinant mouse interferon γ (IFN-γ), oxidized low-density lipoprotein (oxLDL), dihydroethidium (DHE), DiI-oxidized low-density lipoprotein (DiI-oxLDL), hematoxylin-eosin (H & E) stain kit, modified Masson's trichrome stain kit, and modified Oil Red O stain kit were obtained from Beijing Solarbio Science & Technology Co., Ltd. Cy5-baicalein was purchased from Xi'an Qiyue Biology.

Techniques: Fluorescence, Staining, Expressing

Journal: Bioactive Materials

Article Title: Carrier free oral Co-delivery of atorvastatin via baicalein-copper-network for atherosclerosis therapy through senescence reversal and multi-mechanistic synergy

doi: 10.1016/j.bioactmat.2025.12.036

Figure Lengend Snippet: Macrophage reprogramming ability of CMA. (A) Representative optical images of RAW264.7. Scale bar = 50 μm. (B) Confocal laser scanning microscopy images of RAW264.7 cells stained with CD206 antibody, with nuclei counterstained with DAPI. Scale bar = 50 μm. (C) Quantitative analysis of mean CD206 fluorescence intensity. (D) Flow cytometric analysis of CD206 expression in RAW264.7 treated with BAI, Cu-PBS, Cu-MON, ATV, and CMA for 24 h. (E) Relative expression levels of Arg-1, VEGF, TNF-α, and IL-1β in cell supernatants. (F) Flow cytometry scatter plots of iNOS and CD206 expression in RAW264.7 pretreated with LPS + IFN-γ for 24 h followed by treatment with BAI, Cu-PBS, Cu-MON, ATV, and CMA for 24 h. (G–I) Flow cytometric analysis of M1/M2 expression (MFI ratio), iNOs expression, CD206 expression in RAW264.7. (J) Relative expression levels of TGF-β, Arg-1, VEGF, TNF-α, and IL-1β in supernatants from cells treated as in F. Data represent mean ± SD (n = 3–6 independent experiments). Statistical significance: ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001 versus Control; ns = not significant.

Article Snippet: Baicalein (BAI), copper chloride (CuCl 2 ·2H 2 O), and Atorvastatin (ATV) were purchased from Macklin Inc. Lipopolysaccharides (LPS), recombinant mouse interferon γ (IFN-γ), oxidized low-density lipoprotein (oxLDL), dihydroethidium (DHE), DiI-oxidized low-density lipoprotein (DiI-oxLDL), hematoxylin-eosin (H & E) stain kit, modified Masson's trichrome stain kit, and modified Oil Red O stain kit were obtained from Beijing Solarbio Science & Technology Co., Ltd. Cy5-baicalein was purchased from Xi'an Qiyue Biology.

Techniques: Confocal Laser Scanning Microscopy, Staining, Fluorescence, Expressing, Flow Cytometry, Control

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: KSR2 functions as a metabolic checkpoint for anti-PD-1 resistance by reprogramming glucose metabolism

doi: 10.1007/s00262-026-04394-z

Figure Lengend Snippet: KSR2 overexpression associated with an immunosuppressive tumor microenvironment. A Flow cytometry analysis of tumor-infiltrating CD4⁺ T, CD8⁺ T, and regulatory T cells ( n = 3 mice). B , C ELISA quantification of GzmB ( B ) and IFN-γ ( C ) levels ( n = 3 mice). D MIF analysis (CD8, red; GzmB, green; Treg, yellow; DAPI, blue) and cell quantification (3 fields/sample; n = 3 mice). Mean ± SD. two-tailed unpaired t test or Welch’s t test (* P < 0.05, ** P < 0.01, *** P < 0.001)

Article Snippet: Mouse tumor homogenates were prepared, and concentrations of granzyme B and IFN-γ were measured using the mouse granzyme B ELISA kit (E-EL-M0594, Elabscience) and mouse IFN-γ ELISA kit (E-HSEL-M0007, Elabscience), respectively, according to the manufacturers’ protocols.

Techniques: Over Expression, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: KSR2 functions as a metabolic checkpoint for anti-PD-1 resistance by reprogramming glucose metabolism

doi: 10.1007/s00262-026-04394-z

Figure Lengend Snippet: KSR2 knockdown reverses metabolic reprogramming and reinstates anti-tumor immunity. A Differential metabolite abundance in control versus Ksr2-knockdown tumors ( n = 3 mice). B IHC analysis of LDHA and HK2 expression. 3 fields/sample; n = 3 mice. Scale bars, 100μm, 20 μ m. C , D GzmB ( C ) and IFN-γ ( D ) levels in tumors by ELISA ( n = 3). E MIF analysis and cell quantification (3 fields/sample; n = 3 mice). Mean ± SD. Two-tailed unpaired t test or Welch’s t test (* P < 0.05, ** P < 0.01, *** P < 0.001)

Article Snippet: Mouse tumor homogenates were prepared, and concentrations of granzyme B and IFN-γ were measured using the mouse granzyme B ELISA kit (E-EL-M0594, Elabscience) and mouse IFN-γ ELISA kit (E-HSEL-M0007, Elabscience), respectively, according to the manufacturers’ protocols.

Techniques: Knockdown, Control, Expressing, Enzyme-linked Immunosorbent Assay, Two Tailed Test