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: Spatial heterogeneity of immune cells in the tumor microenvironment. Immunophenotypic profiling across invasion states: Non-invaded pseudocapsule (n=9; pathologically tumor-free), tumor-invaded pseudocapsule (n=23), control dura mater from non-invasive tumor cases (n=10), tumor-invaded dura mater (n=21), tumor-invaded mucosa (n=10) and non-invaded mucosa (n=53). (A) Representative immunohistochemistry images for the detection of macrophages (IBA-1 + ), CD4 + T cells, CD8 + T cells and CD19 + B cells. Arrows indicate CD19-positive cells. (B) Quantification of macrophage burden (IBA-1 + immunoreactive area; %). (C) Quantification of CD4 + T-cell density (cells per HPF). (D) Quantification of CD8 + T-cell density (cells per HPF). (E-J) Spatial heterogeneity of macrophage phenotypes. (E) Multiplex immunofluorescence images showing IBA-1 + (red), HLA-DR + (green; M1-like) and CD206 + (magenta; M2-like) macrophage distributions at the IF of the pseudocapsule, dura mater and mucosa, and in non-invaded mucosa. (F) Grayscale-intensity distributions for IBA-1 quantified using ImageJ. (G) M1 immunoreactive area (% of microscopic field) in each group (TIM, TIM-IF, TIDM, TIDM-IF, TIP and TIP-IF). (H) M2 immunoreactive area (% of microscopic field) in each group (TIM, TIM-IF, TIDM, TIDM-IF, TIP and TIP-IF). (I) M1 immunoreactive area (% of microscopic field) in the TIM and NIM groups. (J) M2 immunoreactive area (% of microscopic field) in the TIM and NIM groups. (B-D) Kruskal-Wallis test with prespecified Dunn's post hoc planned comparisons (NIM vs. TIM/NIP/TIP/NIDM/TIDM) and Bonferroni correction. (G and H) Unpaired comparisons among TIM, TIDM and TIP, and separately among TIM-IF, TIDM-IF and TIP-IF, were performed using the Kruskal-Wallis test followed by Dunn's multiple-comparisons test, whereas paired comparisons between each tumor region and its matched IF region were performed using the two-tailed Wilcoxon signed-rank test. Bonferroni correction was applied across all nine comparisons performed in this analysis. (I and J) Unpaired comparisons were analyzed using a two-tailed Mann-Whitney U test. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. HPF, high-power field; IBA-1, ionised calcium binding adaptor molecule 1; IF, invasive front; NIDM, non-invaded dura mater; NIM, non-invaded mucosa; NIP, non-invaded pseudocapsule; ns, not significant; TIDM, tumor-invaded dura mater; TIM, tumor-invaded mucosa; TIP, tumor-invaded pseudocapsule.

Article Snippet: RAW264.7 macrophages in the M0, M1 or M2 state were treated with IgG (10 μg/ml; cat. no. 14-4714-85; Invitrogen; Thermo Fisher Scientific, Inc.) or anti-CD47 monoclonal antibody (mAb) (10 μg/ml; cat. no. 16-0479-85; Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C for 12 h. For polarization, M1 macrophages were induced with lipopolysaccharide (100 ng/ml; cat. no. HY-D1056; MedChemExpress) plus IFN-γ (20 ng/ml; cat. no. RP01070; ABclonal Biotech Co., Ltd.) for 24 h at 37°C, whereas M2 macrophages were induced with IL-4 (20 ng/ml; cat. no. RP01161; ABclonal Biotech Co., Ltd.) for 24 h at 37°C.

Techniques: Control, Immunohistochemistry, Multiplex Assay, Immunofluorescence, Two Tailed Test, MANN-WHITNEY, 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: RAW264.7 macrophages in the M0, M1 or M2 state were treated with IgG (10 μg/ml; cat. no. 14-4714-85; Invitrogen; Thermo Fisher Scientific, Inc.) or anti-CD47 monoclonal antibody (mAb) (10 μg/ml; cat. no. 16-0479-85; Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C for 12 h. For polarization, M1 macrophages were induced with lipopolysaccharide (100 ng/ml; cat. no. HY-D1056; MedChemExpress) plus IFN-γ (20 ng/ml; cat. no. RP01070; ABclonal Biotech Co., Ltd.) for 24 h at 37°C, whereas M2 macrophages were induced with IL-4 (20 ng/ml; cat. no. RP01161; ABclonal Biotech Co., Ltd.) for 24 h at 37°C.

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: 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: RAW264.7 macrophages in the M0, M1 or M2 state were treated with IgG (10 μg/ml; cat. no. 14-4714-85; Invitrogen; Thermo Fisher Scientific, Inc.) or anti-CD47 monoclonal antibody (mAb) (10 μg/ml; cat. no. 16-0479-85; Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C for 12 h. For polarization, M1 macrophages were induced with lipopolysaccharide (100 ng/ml; cat. no. HY-D1056; MedChemExpress) plus IFN-γ (20 ng/ml; cat. no. RP01070; ABclonal Biotech Co., Ltd.) for 24 h at 37°C, whereas M2 macrophages were induced with IL-4 (20 ng/ml; cat. no. RP01161; ABclonal Biotech Co., Ltd.) for 24 h at 37°C.

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

Journal: Skeletal Muscle

Article Title: TRPV1 manipulating polarization of M1/M2 macrophages to promote skeletal muscle regeneration

doi: 10.1186/s13395-026-00417-6

Figure Lengend Snippet: TRPV1 activation leads to a difference in macrophage infiltration with a remarkable increase of M1 and a decrease of M2 in number in vivo. A Representative immunofluorescence images of F4/80 after treatment with CAP and CPZ in CTX-induced muscle tissue at 4d. B The percentage of F4/80-positive cells between four groups at 4d post-injury ( n = 5 animals per experimental group; mean ± SD; One-way ANOVA). C , D Representative immunofluorescence images showed the relative fluorescence intensity of TRPV1 in these F4/80 + macrophages in each group ( n = 5 animals per experimental group; mean ± SD; One-way ANOVA). E , F Representative immunofluorescence images showed the infiltration of numbers of M1 (F4/80 + and CD86+) and M2 (F4/80 + and CD206+) macrophages in different groups at different intervals. G , H The proportion of M1 (F4/80 + and CD86+) macrophages and M2 macrophages (F4/80 + and CD206+) between four groups at 2-8days post-injury ( n = 5 animals per experimental group; mean ± SD; Two-way ANOVA). Statistical significance was set at P < 0.05. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Scale bar, 50 μm

Article Snippet: Macrophage subtypes (M1: CD86-positive and F4/80-positive; M2: CD206-positive and F4/80-positive) were counted independently by two pathologists in five randomly selected non-overlapping fields per section at 400× magnification using a fluorescence microscopy (ECLIPSE Ti2, Nikon, Japan).

Techniques: Activation Assay, In Vivo, Immunofluorescence, Fluorescence

Journal: Skeletal Muscle

Article Title: TRPV1 manipulating polarization of M1/M2 macrophages to promote skeletal muscle regeneration

doi: 10.1186/s13395-026-00417-6

Figure Lengend Snippet: TRPV1 regulates M1/M2 macrophage polarization to promote myogenic differentiation in C2C12 cells. A C2C12 myoblasts were co-cultured with M1 or M2 macrophages for 4 days via a transwell cell culture insert. B Representative Western blot bands of MyoD and MYH3 in C2C12 myoblasts after being co-cultured with M1 or M2 macrophages for 4 days. C , D The protein expression of MyoD and MYH3 in those C2C12 myoblasts which were co-cultured with M1or M2 macrophages after CAP and CPZ treatment ( n = 3 independent replicates in cells; mean ± SD; Two-way ANOVA). E , F Representative immunofluorescence images showed myotubes fusion index in C2C12 cells ( n = 5 independent random fields of cells; mean ± SD; Two-way ANOVA). Statistical significance was set at P < 0.05. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Scale bar, 50 μm

Article Snippet: Macrophage subtypes (M1: CD86-positive and F4/80-positive; M2: CD206-positive and F4/80-positive) were counted independently by two pathologists in five randomly selected non-overlapping fields per section at 400× magnification using a fluorescence microscopy (ECLIPSE Ti2, Nikon, Japan).

Techniques: Cell Characterization, Cell Culture, Western Blot, Expressing, Immunofluorescence