Journal: Molecular Cancer

Article Title: Integrated multi-omics identifies a CD54 + iCAF-ITGAL + macrophage niche driving immunosuppression via CXCL8-PDL1 axis in cervical cancer

doi: 10.1186/s12943-025-02471-y

Figure Lengend Snippet: CD54⁺ iCAFs promote monocyte migration and M2-like polarization through CCL2 secretion. A Left: Transwell migration assay of THP-1 cells co-cultured with CD54⁺ iCAFs or normal fibroblasts (NFs) for 48 h. Right: Quantification of migrated cells (n = 3 independent experiments). Scale bar: 100 μm. B-C THP-1 cells co-cultured with CD54⁺ iCAFs show upregulated expression of M2-like macrophage biomarkers and cytokines. D CD54⁺ iCAFs were transfected with CD54-targeting siRNA (si-CD54) or negative control siRNA (si-NC). Volcano plot of differentially expressed genes (DEGs) identified by RNA-seq analysis between si-CD54 and si-NC groups. E CD54 + iCAFs were transfected with si-CD54 or si-NC. Then, CD54 and CCL2 protein levels were assessed by Western blot. Left: Representative images. Right: Quantification from n = 3 independent experiments. F ELISA quantification of CCL2 secretion from CD54⁺ iCAFs and NFs. G Left: THP-1 cell migration in response to recombinant CCL2 treatment (50 ng/ml, 48 h) or PBS (Control group). Right: Quantification of migrated cells. All quantitative data are presented as mean ± SD. Statistical significance was determined using Student’s t-test for panels A (right), B, C, E (right), F, and G (right). *: P < 0.05, **: P < 0.01, ***: P < 0.001; ns, not significant

Article Snippet: A neutralizing anti-human CCL2 antibody (MedChemExpress) or an IgG isotype control was introduced to the culture medium.

Techniques: Migration, Transwell Migration Assay, Cell Culture, Expressing, Transfection, Negative Control, RNA Sequencing, Western Blot, Enzyme-linked Immunosorbent Assay, Recombinant, Control

Journal: Molecular Cancer

Article Title: Integrated multi-omics identifies a CD54 + iCAF-ITGAL + macrophage niche driving immunosuppression via CXCL8-PDL1 axis in cervical cancer

doi: 10.1186/s12943-025-02471-y

Figure Lengend Snippet: CD54⁺ iCAF-driven macrophage reprogramming promotes CXCL8 expression and tumor progression in vivo. A THP-1-derived macrophages, transfected with control siRNA (Control group) or ITGAL-targeting siRNA (Treatment group), were co-cultured with CD54⁺ iCAFs for 48 h and subsequently subjected to RNA-seq analysis. Volcano plot shows differentially expressed genes (DEGs) between control and treatment groups. B Left: Representative flow cytometry plots of CD54⁺ iCAF abundance. Middle: Quantitative analysis of CXCL8⁺ cell frequencies in CD54⁺ iCAF-high tumors (n = 3). Right: quantitative analysis of CXCL8 expression in CD54 + iCAF-high versus CD54 + iCAF-low (Displaying in Supplementary Fig. 5F) tumors. C Flow cytometric quantification of CXCL8-producing immune cell subsets in cervical cancer tissues. D Multiplex immunohistochemistry images showing macrophage-specific CXCL8 expression in cervical cancer tissue (Scale bar: 100 μm). E NIH/3T3 fibroblasts transfected with CD54 overexpression plasmid (OE-CD54) or empty vector control (OE-NC). CD54 and CCL2 secretion levels were quantified by ELISA. F Representative tumor images from C57BL/6 mice co-injected with TC-1 cells and NIH/3T3 fibroblasts expressing CD54 or empty vector (Control). G Tumor growth curves measured every 3 days (n = 3 mice/group). H-I Flow cytometry analysis of CD206⁺ macrophage infiltration in tumors from (F). I (Right): Quantification of CD206⁺ macrophage proportions. J Serum MIP-2 levels measured by ELISA in experimental groups from (F). All quantitative data are presented as mean ± SD. Statistical significance was determined using Student’s t-test for panels B (right), E, G(right), I (right), and J. *: P < 0.05, **: P < 0.001, ***: P <0.001

Article Snippet: A neutralizing anti-human CCL2 antibody (MedChemExpress) or an IgG isotype control was introduced to the culture medium.

Techniques: Expressing, In Vivo, Derivative Assay, Transfection, Control, Cell Culture, RNA Sequencing, Flow Cytometry, Multiplex Assay, Immunohistochemistry, Over Expression, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Injection

Journal: Molecular Cancer

Article Title: Integrated multi-omics identifies a CD54 + iCAF-ITGAL + macrophage niche driving immunosuppression via CXCL8-PDL1 axis in cervical cancer

doi: 10.1186/s12943-025-02471-y

Figure Lengend Snippet: CXCL8 correlates with CD8⁺ T cell exclusion and promotes PD-L1 expression on macrophages through cell-contact and soluble-factor dependent mechanisms. A CIBERSORT analysis showing an inverse correlation between CXCL8 mRNA levels and CD8⁺ T cell infiltration. B Representative immunohistochemistry (IHC) images showing CD8⁺ T cell density in high- versus low-CXCL8 expressing tumors (Scale bar: 100 μm). C Negative correlation between protein levels of CXCL8 and PD-L1 based on IHC scoring (Pearson correlation). D Left: Flow cytometry plots of PD-L1⁺ cells. Right: Quantification of PD-L1 expression in high- versus low-CXCL8 tumors (n = 3 per group). E Representative IHC staining confirming macrophage-specific PD-L1 expression (Scale bar: 100 μm). F Left: Gating strategy for identifying PD-L1⁺ cells. Right: Quantification of PD-L1 expression across immune cell subtypes, showing macrophage dominance (n = 6). G Left: Flow cytometry profiles of PD-L1 expression. Right: PD-L1 levels in high- versus low-CXCL8 tumors. H Flow cytometry analysis of CXCL8 and PD-L1 expression on CD68+ macrophages co-cultured with CD54⁺ iCAFs under direct contact or Transwell conditions, with normal fibroblasts (NFs) and macrophage-only cultures as controls. The bar graph shows geometric mean fluorescence intensity from three independent experiments. Corresponding representative flow cytometry plots are shown in Supplementary Fig. 6D. I CXCL8 and PD-L1 expression on macrophages after co-culture with CD54⁺ iCAFs and treatment with IgG control, anti-CD54, or anti-ITGAL blocking antibodies. See Supplementary Fig. 6E for flow plots. J CXCL8 and PD-L1 expression on macrophages transfected with control siRNA (si-NC) or ITGAL-targeting siRNA (si-ITGAL), with or without CD54 + iCAF co-culture. Corresponding representative flow cytometry plots are shown in Supplementary Fig. 6F. K CXCL8 and PD-L1 expression on macrophages co-cultured with CD54⁺ iCAFs and treated with IgG control or anti-CCL2 neutralizing antibody. Representative flow plots are provided in Supplementary Fig. 6G. Data are presented as mean ± SD. Statistical tests used: two-tailed Student’s t-test (D, right; G, right; K); one-way ANOVA with Tukey's multiple comparisons test (F, right; H, I, J). **: P < 0.01, ***: P < 0.001; ns, not significant

Article Snippet: A neutralizing anti-human CCL2 antibody (MedChemExpress) or an IgG isotype control was introduced to the culture medium.

Techniques: Expressing, Immunohistochemistry, Flow Cytometry, Cell Culture, Fluorescence, Co-Culture Assay, Control, Blocking Assay, Transfection, Two Tailed Test

Journal: Science Advances

Article Title: Immunosuppressive macrophages determine the effect of cellular senescence on tumor progression

doi: 10.1126/sciadv.adx2988

Figure Lengend Snippet: ( A ) Tumor samples from mice treated with vehicle or AP21967 at 9 weeks were homogenized, and the levels of the indicated cytokines were analyzed using a cytokine array. The log 2 FC (AP21967/vehicle) for each cytokine is presented. ENA-78, epithelial-derived neutrophil-activating peptide 78; GROα, growth-regulated oncogene-alpha. ( B ) Levels of CCL2 and CXCL1 were quantified by ELISA from the same tumor homogenates as in (A). ( C ) UMAP plots showing the expression of the most up-regulated cytokines (CCL2, CCL3, and CCL7) and their receptors (CCR1, CCR2, and CCR5) in different cell populations. Relevant cell clusters are indicated. N, neutrophils; MØ, macrophages; E, endothelial cells; F, fibroblasts. ( D ) Flow cytometry analysis of myeloid cells purified from tumors obtained from SuSe mice treated with vehicle or AP21967. We identified TAMs and cells positive for the myeloid marker CD11B and for the macrophage marker F4/80. ( E ) The same cells as in (D) were cultured for 24 hours—in the presence of vehicle or anti-CCL2—and the presence of CCL2 in the conditioned medium was determined by ELISA. ( F ) Schematic representation of the specificities of receptors for CCL2, CCL3, and CCL7. ( G ) Flow cytometry analysis of CCR2 in the same cells as in (D). FSH-H, forward scatter height.

Article Snippet: For selective CCL2 blocking, 4-week-old PyMT/SuSe mice were treated every 3 days with anti-CCL2 antibody (#BE0185, BioXCell,) at a concentration of 2 μg/g of body weight.

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Expressing, Flow Cytometry, Purification, Marker, Cell Culture

Journal: Molecular human reproduction

Article Title: Regulated C-C motif ligand 2 (CCL2) in luteal cells contributes to macrophage infiltration into the human corpus luteum during luteolysis.

doi: 10.1093/molehr/gav028

Figure Lengend Snippet: Figure 1 Expression of CCL2 in the human CL is associated with the number of infiltrating macrophages. The CD68-positive infiltrating macrophages are distributed at the border of granulosa-lutein (g) and theca-lutein cells (t) and dispersed in the CL parenchyma at mid-luteal (ML) phase (A). CD68-positive macrophages tend to increase in the CL at late-luteal (LL) phase (B). Numerous CD68-positive cells are found in the CL collected during menstruation (Men), and the number of CD68-positive macrophages is significantly increased in the regressing CL at this stage (C and D). CCL2 protein is localized in the cytoplasm of granulosa-lutein and theca-lutein cells in the human CL at ML phase (E). Endothelial cells are weakly immunoreactive for CCL2 (arrows in E) while non-steroidogenic cells distributed in the CL parenchyma are negative in reaction (asterisks in the insert of E). The mRNA expression ofCCL2peaksintheCLcollected duringmenstruation(F) andisreducedby invivoadministrationofhCG(G).EL,early-lutealphase; R,theCLrescuedfrom luteolysis by in vivo administration of hCG. *P , 0.05, **P , 0.01.

Article Snippet: After pretreatment with normal goat serum for 60 min at room temperature, they were incubated with mouse anti-human CD68 (1:100; PG-M1, Dako UK Ltd, Cambridgeshire, UK), mouse anti-human CCL2 (1:100; MCA2486, AbD serotec, Raleigh, NC, USA), rabbit anti-3b-HSD (1:1000; kind gift from Prof. Ian Mason, The University of Edinburgh) or rabbit anti-human prostaglandin E synthase (PGES; 1:250; 160140, Cayman Chemical, Ann Arbor, MI, USA) in goat serum at 48C overnight.

Techniques: Expressing, In Vivo

Journal: Molecular human reproduction

Article Title: Regulated C-C motif ligand 2 (CCL2) in luteal cells contributes to macrophage infiltration into the human corpus luteum during luteolysis.

doi: 10.1093/molehr/gav028

Figure Lengend Snippet: Figure 2 Regulation of CCL2 mRNA expression in cultured LGCs. The mRNA expression of CCL2 is significantly decreased by the treatment of hCG in vitro (A). Withdrawal of hCG significantly enhances the CCL2 expression in LGCs on Day 8 and 9 (B). Luteotrophic PGE decreases the expression of CCL2 (C),whereasluteolytic PGFpromotesitsexpression(D).Amacrophagecytokine,TNF,issignificantlyincreasedintheCLcollectedduringmenstruation(E) and its expression is positively correlated to the expression of CCL2 in the human CL (F). The expression of TNF is reduced in the rescued CL by in vivo administrationof hCG (G). CCL2 expression is promotedby TNF treatment in LGCs in vitro(H). PKC activator(PMA) significantlyenhancesthe expression ofCCL2(I)whileinhibitorsforPI3kinase(LY294002:LY)andPKA(H89)suppressitsexpression(JandK).Cont,control;DMSO,dimethylsulfoxideusedas a vehicle for PMA, LY and H89. EL, early-luteal; EtOH, ethanol used as avehicle for prostaglandins; LL, late-luteal phase; Men, menstruation; ML, mid-luteal phase; R, rescued CL by in vivo administration of hCG. *P , 0.05, **P , 0.01, ****P , 0.0001.

Article Snippet: After pretreatment with normal goat serum for 60 min at room temperature, they were incubated with mouse anti-human CD68 (1:100; PG-M1, Dako UK Ltd, Cambridgeshire, UK), mouse anti-human CCL2 (1:100; MCA2486, AbD serotec, Raleigh, NC, USA), rabbit anti-3b-HSD (1:1000; kind gift from Prof. Ian Mason, The University of Edinburgh) or rabbit anti-human prostaglandin E synthase (PGES; 1:250; 160140, Cayman Chemical, Ann Arbor, MI, USA) in goat serum at 48C overnight.

Techniques: Expressing, Cell Culture, In Vitro, In Vivo, Control

Journal: Molecular human reproduction

Article Title: Regulated C-C motif ligand 2 (CCL2) in luteal cells contributes to macrophage infiltration into the human corpus luteum during luteolysis.

doi: 10.1093/molehr/gav028

Figure Lengend Snippet: Figure 5 Relationship between the function of luteal cells and macrophage infiltration. Two serial sections of the human CL at the late-luteal phase were immunostained for a PGES (A) or a macrophage marker (CD68) (B). Healthy luteal cells contained abundant immunoreactivity for PGES (arrow in A) and there are fewCD68-positive macrophages around them (arrow in B). Abundant CD68-positive cells are found in the region with PGES-negative granulosa- lutein cells (asterisks in A and B). The mRNA expression of PGES (C) and HSD3B1 (D) tends to be negatively correlated to that of CCL2 in the human CL.

Article Snippet: After pretreatment with normal goat serum for 60 min at room temperature, they were incubated with mouse anti-human CD68 (1:100; PG-M1, Dako UK Ltd, Cambridgeshire, UK), mouse anti-human CCL2 (1:100; MCA2486, AbD serotec, Raleigh, NC, USA), rabbit anti-3b-HSD (1:1000; kind gift from Prof. Ian Mason, The University of Edinburgh) or rabbit anti-human prostaglandin E synthase (PGES; 1:250; 160140, Cayman Chemical, Ann Arbor, MI, USA) in goat serum at 48C overnight.

Techniques: Marker, Expressing