Journal: Frontiers in Immunology

Article Title: A novel Bacillus megaterium /ω-3 PUFA synbiotic attenuates colon inflammation and promotes a pro-resolving macrophage profile ex-vivo

doi: 10.3389/fimmu.2026.1916994

Figure Lengend Snippet: Representative images of immunofluorescence staining with CD68 and CD163 of human colon sections obtained from healthy patients in basal condition (A) , treated for 24 hours with 2,5 μL of SynΩ3 formulation (B) , or with ω-3 lysine salt dispersion alone (C) . DAPI-stained nuclei are in blue. Scale bar: 100μm; zoom inset up: 50μm; zoom inset down: 50μm. (D) The bar graph shows the CD68 and CD163 co-localization rate of human colon sections obtained from IBD patients in basal condition treated for 24 hours with 2,5 μL of SynΩ3 formulation or with ω-3 lysine salt dispersion alone. n= 2.

Article Snippet: Colon sections were incubated overnight at 4 °C with the following primary antibodies: mouse monoclonal anti-CD68 antibody (1:50, Abcam, catalog ab955, clone KP1), rabbit polyclonal anti-CD163 antibody (1:50 ABclonal, catalog A8383), and mouse monoclonal anti-CD206 antibody (1:50, Proteintech, catalog 60143-1).

Techniques: Immunofluorescence, Staining, Formulation, Dispersion

Journal: Frontiers in Immunology

Article Title: A novel Bacillus megaterium /ω-3 PUFA synbiotic attenuates colon inflammation and promotes a pro-resolving macrophage profile ex-vivo

doi: 10.3389/fimmu.2026.1916994

Figure Lengend Snippet: Representative images of immunofluorescence staining with CD68 and CD163 of human colon sections obtained from UC patients in basal condition (A) , treated for 24 hours with 2,5 μL of SynΩ3 formulation (B) or with ω-3 lysine salt dispersion alone (C) . DAPI-stained nuclei are in blue. Scale bar: 100μm; zoom inset up: 50μm; zoom inset down: 50μm. (D) The bar graph shows the CD68 and CD163 co-localization rate of human colon sections obtained from UC patients in basal condition treated for 24 hours with 2,5 μL of SPMs generated by SynΩ3 formulation or with ω-3 lysine salt dispersion alone. n= 3.

Article Snippet: Colon sections were incubated overnight at 4 °C with the following primary antibodies: mouse monoclonal anti-CD68 antibody (1:50, Abcam, catalog ab955, clone KP1), rabbit polyclonal anti-CD163 antibody (1:50 ABclonal, catalog A8383), and mouse monoclonal anti-CD206 antibody (1:50, Proteintech, catalog 60143-1).

Techniques: Immunofluorescence, Staining, Formulation, Dispersion, Generated

Journal: Bioactive Materials

Article Title: Immunomodulatory effects of biodegradable Mg–Cu–Zn alloy in esophageal cancer

doi: 10.1016/j.bioactmat.2026.02.046

Figure Lengend Snippet: Distribution of CD163 + M2 TAMs in AKR-derived allograft tumor tissues from immunocompetent C57BL/6 mice. (a, b) Representative IHC staining images showing CD163 + M2 TAMs in the (a) peritumoral stroma and (b) tumor islets. Lower panels display higher-magnification views of the regions outlined by red dashed boxes. (c, d) Quantification of CD163 + cells in the (c) peritumoral stroma and (d) tumor islets. (e) Comparison of CD163 + cell density between the peritumoral stroma and tumor islets. (f) Total number of CD163 + cells in allograft tumors (peritumoral stroma and tumor islets combined). (g, h) Comparison of the density between iNOS + cells and CD163 + cells in the (g) peritumoral stroma and (h) tumor islets. (i, j) Quantification of iNOS + /CD163 + ratio in the (i) peritumoral stroma and (j) tumor islets. p < 0.05 (∗), p < 0.01 (∗∗), p < 0.001 (∗∗∗). A field of view is ∼0.086 mm 2 in (c−j).

Article Snippet: Tissue sections were then incubated with primary antibodies against iNOS (22226-1-AP, ProteinTech, China), CD163 (A26411PM, Abclone, China), CD8 (SP16, Maixin, China), CD4 (SP35, Maixin, China) or Ki-67 (12202S, Cell Signaling Technology) for 12 h at 4 °C, followed by secondary antibodies (Beyotime Biotechnology, Nantong, China).

Techniques: Derivative Assay, Immunohistochemistry, Comparison

Journal: Frontiers in Immunology

Article Title: Identification of LILRB4 as a regulator of M2c macrophages and a potential immunotherapeutic target in ovarian cancer

doi: 10.3389/fimmu.2026.1866073

Figure Lengend Snippet: LILRB4 regulates M2c macrophage polarization. (A) Patients in the TCGA-OV cohort were stratified into high- and low-LILRB4 expression groups to assess differences in M2 macrophage infiltration. (B) LILRB4 expression levels in M0, M1, and M2 macrophages based on the GEPIA database. (C) TCGA-OV samples were categorized into high and low M2 macrophage infiltration groups. OS was analyzed based on LILRB4 expression using the TIMER database; statistical significance was determined by the log-rank test. (D) Correlation analysis LILRB4 and CD206 (M2a), HLA-DR (M2b), CD163 (M2c), CD14 (M2d) by GSE12470 cohort. (E) Correlation analysis between LILRB4 expression and the M2c markers CD163 in the TCGA-OV cohort. (F, H) Flow cytometry analysis of CD163 expression in macrophages following co-culture with OVCAR8 (F) or ES2 (H) cells with NC or LILRB4 knockdown. (G, I) qRT-PCR analysis of IL-10 and TGFβ1 mRNA expression in macrophages co-cultured with OVCAR8 (G) or ES2 (I) cells under control (NC) or LILRB4 knockdown conditions. (J) Flow cytometry analysis of CD163 expression in macrophages following co-culture with HEY cells with NC or OE-LILRB4. (K) A schematic diagram showing the process of collecting tumor cell CM and co-culturing it with macrophages for 48 hours before conducting flow cytometry detection. (L) Macrophages were treated with CM from LILRB4-overexpressing tumor cells or control tumor cells, and the proportion of CD163-positive macrophages was assessed (left) and quantitatively compared between the 2 groups (right). (M) Macrophages were treated with CM from LILRB4-overexpressing tumor cells or control tumor cells, and the expression levels of IL-10 and TGFβ1 in macrophages were measured by qRT-PCR. Data are presented as mean ± SD. Statistical analysis was performed using unpaired two-tailed Student’s t-test (A, F-J, L, M). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: All antibodies used in this study were commercially available: LILRB4 (A7073; ABclonal, Wuhan, China; 1:2000 for WB and 1:200 for immunohistochemistry), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (AB2100; NCM Biotech, Suzhou, China; 1:20,000), F4/80 (A27257; ABclonal, Wuhan, China; 1:200), Ki67 (A20018; ABclonal, Wuhan, China; 1:200), CD68 (A24386PM; ABclonal, Wuhan, China; 1:200), phospho-NF-κB (p-NF-κB; TP56372; Abmart, Shanghai, China; 1:1000), CD163 (A8383SP; ABclonal, Wuhan, China; 1:200), CD8 (A23081; ABclonal, Wuhan, China; 1:200), granzyme B (GZMB) (A2557; ABclonal, Wuhan, China; 1:200), and goat anti-rabbit IgG (H+L)-HRP conjugate (LF102; Epizyme Biotech, Shanghai, China; 1:3000).

Techniques: Expressing, Flow Cytometry, Co-Culture Assay, Knockdown, Quantitative RT-PCR, Cell Culture, Control, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Identification of LILRB4 as a regulator of M2c macrophages and a potential immunotherapeutic target in ovarian cancer

doi: 10.3389/fimmu.2026.1866073

Figure Lengend Snippet: LILRB4 promotes M2c macrophage differentiation via CCL5/NF-κB signaling. (A) In the OVCAR8 cells, qRT-PCR was used to analyze the mRNA expression levels of CCL2, CCL5, CCL18, CCL22, CXCL12 and CXCL8. (B) Correlation analysis between CCL5 (left) and CCL22 (right) expression and M2 macrophage infiltration based on the TIMER database. (C) Expression levels of the M2c macrophage marker CD163 in macrophages following treatment with CCL5 or CCL22. (D) The expression of CCL5 mRNA in ES2 cells either under normal control conditions or under sh-LILRB4 treatment was analyzed by qRT-PCR. (E) The expression of CCL5 mRNA in HEY cells under different treatment was analyzed by qRT-PCR. (F) qRT-PCR analysis of CCL5 mRNA expression in ovarian tumor tissues from the indicated groups. (G) Correlation analysis between LILRB4 expression and CCL5 in the TCGA-OV cohort. (H) Flow cytometry analysis of CD11b + CD163 + macrophages in the control and CCL5-treated groups (left), with corresponding quantification shown on the right. (I) Correlation analysis between CCL5 expression and key signaling molecules (NFKB1, AKT1, mTOR, STAT6, MAPK1, and NOTCH1) in the TCGA-OV cohort. (J) WB analysis showing p-p65 levels in macrophages treated with CM from control or LILRB4-overexpressing tumor cells or maraviroc, indicating activation of the NF-κB signaling pathway. (K) Expression levels of CD163 in macrophages under the above treatment conditions (up), with quantitative analysis of the different groups (down). (L) Assessment of p-p65 activation in macrophages following CCL5 treatment. (M) qRT-PCR analysis of CD163, IL-10, and TGFβ1 mRNA expression in macrophages treated with control (NC) or CCL5 or CCL5 with PDTC. Data are presented as mean ± SD. Statistical analysis was performed with unpaired two-tailed Student’s t-test (A, C-F, H) or unpaired two-tailed Student’s t-test (K, M). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: All antibodies used in this study were commercially available: LILRB4 (A7073; ABclonal, Wuhan, China; 1:2000 for WB and 1:200 for immunohistochemistry), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (AB2100; NCM Biotech, Suzhou, China; 1:20,000), F4/80 (A27257; ABclonal, Wuhan, China; 1:200), Ki67 (A20018; ABclonal, Wuhan, China; 1:200), CD68 (A24386PM; ABclonal, Wuhan, China; 1:200), phospho-NF-κB (p-NF-κB; TP56372; Abmart, Shanghai, China; 1:1000), CD163 (A8383SP; ABclonal, Wuhan, China; 1:200), CD8 (A23081; ABclonal, Wuhan, China; 1:200), granzyme B (GZMB) (A2557; ABclonal, Wuhan, China; 1:200), and goat anti-rabbit IgG (H+L)-HRP conjugate (LF102; Epizyme Biotech, Shanghai, China; 1:3000).

Techniques: Quantitative RT-PCR, Expressing, Marker, Control, Flow Cytometry, Activation Assay, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Identification of LILRB4 as a regulator of M2c macrophages and a potential immunotherapeutic target in ovarian cancer

doi: 10.3389/fimmu.2026.1866073

Figure Lengend Snippet: Clinical significance of LILRB4 expression in OC. (A–B) Differences in LILRB4 expression levels were compared among subgroups with venous invasion (A) and lymphatic invasion (B) in the TCGA-OV cohort. (C–D) Correlation analysis between LILRB4 and immune checkpoint molecules in the TCGA-OV cohort, presented as a chord diagram (C) and a lollipop plot (D) . (E) Kaplan–Meier survival analysis of patients from the immunotherapy cohort (esophageal adenocarcinoma treated with immune checkpoint inhibitor-based therapy). OS (left) and PFS (right) were compared between tumors with high and low LILRB4 expression. Patients were stratified according to the optimal cutoff determined by the Kaplan–Meier Plotter platform. Statistical significance was assessed using the log-rank test. (F) Representative images of orthotopic ID8 tumors in C57BL/6 mice (n = 4 per group). (G) Comparison of tumor weight (left) and volume (right) among different groups of C57BL/6 mice (n = 4 per group). (H) IHC analysis of Ki-67 and CD163 expression in tumor tissues (n = 3). All images were captured at the same magnification; scale bar = 100 μm. (I–J) WB analysis of p-p65 expression in tumor tissues (I, n = 3) and corresponding quantitative analysis (J) . (K) IHC analysis of CD8 and GZMB expression in tumor tissues (n = 3). All images were captured at the same magnification; scale bar = 100 μm. (L) A nomogram was constructed based on independent prognostic factors for overall survival. Data are presented as mean ± SD. Statistical analysis was performed using an unpaired two-tailed Student’s t-test. *p < 0.05, **p < 0.01, ****p < 0.0001.

Article Snippet: All antibodies used in this study were commercially available: LILRB4 (A7073; ABclonal, Wuhan, China; 1:2000 for WB and 1:200 for immunohistochemistry), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (AB2100; NCM Biotech, Suzhou, China; 1:20,000), F4/80 (A27257; ABclonal, Wuhan, China; 1:200), Ki67 (A20018; ABclonal, Wuhan, China; 1:200), CD68 (A24386PM; ABclonal, Wuhan, China; 1:200), phospho-NF-κB (p-NF-κB; TP56372; Abmart, Shanghai, China; 1:1000), CD163 (A8383SP; ABclonal, Wuhan, China; 1:200), CD8 (A23081; ABclonal, Wuhan, China; 1:200), granzyme B (GZMB) (A2557; ABclonal, Wuhan, China; 1:200), and goat anti-rabbit IgG (H+L)-HRP conjugate (LF102; Epizyme Biotech, Shanghai, China; 1:3000).

Techniques: Expressing, Comparison, Construct, Two Tailed Test