Journal: Journal for Immunotherapy of Cancer

Article Title: Preventing surgery-induced natural killer cell suppression and metastases by inhibiting PI3K-gamma signaling in myeloid-derived suppressor cells

doi: 10.1136/jitc-2025-013304

Figure Lengend Snippet: Large expansion of phenotypically defined sx-MDSCs comprised mainly of M-MDSC and PMN-MDSCs immediately after surgery (POD1). ( A ) Representative flow plots showing CD33 versus Lineage (CD3/CD56/CD19) for baseline (left) and POD1 (right) PBMCs. ( B ) Representative flow plots gated down on CD33 + Lin − cells showing CD15 versus CD14 expression. ( C ) Healthy controls (n=15), Baseline, and POD1 patients (n=44) proportion of CD33 + Lin − cells (left) and MDSC subsets (right). ( D ) The MFI of HLA-DR gated on M-MDSCs (left) and the proportion of M-MDSCs that are HLA-DR lo (right) in healthy controls, baseline and POD1 patients with cancer. ( E ) Representative histograms of common M-MDSC and PMN-MDSC markers, before and after surgery. Statistical analysis was performed using Wilcoxon matched pairs signed-rank test or Kruskal-Wallis with Dunn’s post test. **** p≤0.0001. FMO, fluorescence minus one control; HLA-DR, human leukocyte antigen - DR isotype; E-MDSC, early stage-MDSC; M-MDSC, monocytic-MDSC; MDSC, myeloid derived suppressor cell; sx-MDSC, surgery-induced MDSC, MFI; mean fluorescence instensity; PBMC, peripheral blood mononuclear cell; PMN-MDSC, polymorphonuclear-MDSC; POD1, postoperative day 1.

Article Snippet: Sx-MDSCs were isolated using the MDSC Isolation Kit (Miltenyi #130-094-538) from surgically stressed mice on POD1.

Techniques: Expressing, Fluorescence, Control, Derivative Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Preventing surgery-induced natural killer cell suppression and metastases by inhibiting PI3K-gamma signaling in myeloid-derived suppressor cells

doi: 10.1136/jitc-2025-013304

Figure Lengend Snippet: Sx-MDSCs suppress NK92-MI cytotoxicity. ( A ) Schematic of MDSC-NK92 co-culture suppression assay with CP450-labeled K562 target cells. ( B ) NK92-MIs were co-cultured at increasing ratios with CD33+ cells for 6 or 24 hours (in the absence of K562s) to assess CD33+ cell death by NK92-MI (left). Sx-MDSCs from POD1 patient samples (n=10) were co-cultured with NK92-MI cells and NK cytotoxicity was measured as % dead K562 (right). ( C ) % suppression was calculated as the reduction of NK cytotoxicity, normalized to NK cells alone (ratio=0). CD33+ cells isolated from baseline and patient-matched POD1 blood were co-cultured with NK92-MI and K562 targets to measure their suppressive capacity (n=5). ( D ) Representative images of sorted cells stained with Giemsa Wright. ( E ) Bulk Sx-MDSCs, M-MDSCs, PMN-MDSCs, and HDN from POD1 patients (n=3) co-cultured with NK92-MI to measure effect on NK cytotoxicity. The NK92-MI alone control group is indicated by the broken horizontal line. Statistical analysis used Kruskal-Wallis with Dunn’s post-test. ****p≤0.0001. HDN, high-density neutrophils; M-MDSC, monocytic-MDSC; MDSC, myeloid-derived suppressor cell; NK, natural killer; sx-MDSC, surgery-induced MDSC, PMN-MDSC, polymorphonuclear-MDSC; POD1, postoperative day 1.

Article Snippet: Sx-MDSCs were isolated using the MDSC Isolation Kit (Miltenyi #130-094-538) from surgically stressed mice on POD1.

Techniques: Co-Culture Assay, Suppression Assay, Labeling, Cell Culture, Isolation, Staining, Control, Derivative Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Preventing surgery-induced natural killer cell suppression and metastases by inhibiting PI3K-gamma signaling in myeloid-derived suppressor cells

doi: 10.1136/jitc-2025-013304

Figure Lengend Snippet: scRNA-seq of cryopreserved PBMCs before and after surgery reveals drastically altered monocyte/myeloid cell expression profiles on POD1. ( A ) Schematic showing PBMCs (cryopreserved) from six matched baseline and POD1 patients were processed for multiplexed scRNA-seq with the 10x Genomics Chromium platform. ( B ) Dot plot displaying the relative expression of the top three marker genes (x-axis) of each cluster (y-axis). ( C ) UMAP plot of scRNA-seq data. Each point corresponds to a single cell and is colored by cluster. ( D ) Identical UMAP embedding as in ( C ), with Baseline and POD1 cells labeled. ( E ) UMAP plot of the CD14+ and CD16+ monocyte population at baseline and POD1. ( F ) GSEA showing the NES of the top upregulated and downregulated pathways. All gene sets are significantly enriched (FDR<0.05). ( G ) UMAP plot showing activity of an M-MDSC gene set from Alshetaiwai et al ; in monocyte populations in Baseline and POD1 samples. ( H ) NMF plots of gene expression programs for NMF 1, 2, 3 and 4 (see for all NMF programs). ( I ) A heatmap of selected genes driving the various NMF gene expression programs (z-score transformed; ranked according to NMF1). FDR, false discovery rate; GSEA, gene set enrichment analysis; M-MDSC, monocytic-myeloid-derived suppressor cell; NES, normalized enrichment scores; NMF, non-negative matrix factorization; PBMC, peripheral blood mononuclear cell; POD1, postoperative day 1; scRNA-seq, single-cell RNA sequencing; UMAP, uniform manifold approximation and projection.

Article Snippet: Sx-MDSCs were isolated using the MDSC Isolation Kit (Miltenyi #130-094-538) from surgically stressed mice on POD1.

Techniques: Expressing, Marker, Labeling, Activity Assay, Gene Expression, Transformation Assay, Derivative Assay, RNA Sequencing

Journal: Journal for Immunotherapy of Cancer

Article Title: Preventing surgery-induced natural killer cell suppression and metastases by inhibiting PI3K-gamma signaling in myeloid-derived suppressor cells

doi: 10.1136/jitc-2025-013304

Figure Lengend Snippet: PI3K inhibitors reverse the suppressive effects of sx-MDSCs on NK cells. ( A ) A library of 147 small molecules covering the major cellular signaling pathways was screened (n=4 experiments) at 1 µM in the MDSC-NK suppression assay. Compounds that improved NK cell cytotoxicity (blue, left y-axis) by >50% from DMSO control (black dotted line), without impacting NK cell viability (red, right y-axis), were considered hits. LY294002 (purple circle) improved cytotoxicity in all screens and was the top hit in 3/4 screens. ( B ) LY294002 improves NKC (n=6). ( C ) Dose response of LY294002 and the effect on NKC and K562 viability. ( D ) pan-PI3K and ( E ) PI3K-γ specific inhibitors (IPI-549, TG100-115) in NK92—MDSC suppression assays. ( F ) pan-PI3K and ( G ) PI3K-γ specific inhibitors (IPI-549, TG100-115) in MDSC suppression assays with primary healthy donor NK cells. Statistical analysis used one-way ANOVA with Dunnett’s multiple comparisons test. ***p<0.0001. ANOVA, analysis of variance; DMSO, dimethyl sulfoxide; MDSC, myeloid-derived suppressor cell; NK, natural killer; sx-MDSC, surgery-induced MDSC.

Article Snippet: Sx-MDSCs were isolated using the MDSC Isolation Kit (Miltenyi #130-094-538) from surgically stressed mice on POD1.

Techniques: Protein-Protein interactions, Suppression Assay, Control, Derivative Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Preventing surgery-induced natural killer cell suppression and metastases by inhibiting PI3K-gamma signaling in myeloid-derived suppressor cells

doi: 10.1136/jitc-2025-013304

Figure Lengend Snippet: PI3K pathway activation in postoperative MDSCs and functional impact of PI3K inhibition. ( A ) Cohort of differentially expressed genes from baseline versus POD1 MDSCs. ( B ) Box plot inference of PI3K pathway activity in baseline versus POD1 MDSCs. ( C ) Pathway diagram of PI3K signaling and Akt phosphorylation. Phosphorylation status of AKT at the T308 (left) and S473 residue (right) (n=5). ( D ) Effect of PI3K inhibitors on pAKT (T308) MFI (top), MDSC-NK suppression (middle), and NKC (bottom). ( E ) Effect of IPI-549 on POD1 expression of anti-inflammatory versus pro-inflammatory mRNA transcripts, normalized to baseline. Statistical analysis used Wilcoxon matched pairs signed-rank tests and one-way ANOVA with Dunnett’s multiple comparisons test (vs DMSO control). ****p>0.0001. ANOVA, analysis of variance; ARG1, arginase-1; CCL, CC chemokine ligand; DMSO, dimethyl sulfoxide; IL, interleukin; MDSC, myeloid-derived suppressor cell; mRNA, messenger RNA; MFI, mean fluorescence intensity; NK, natural killer; POD1, postoperative day 1; TGF, transforming growth factor; TNF, tumor necrosis factor.

Article Snippet: Sx-MDSCs were isolated using the MDSC Isolation Kit (Miltenyi #130-094-538) from surgically stressed mice on POD1.

Techniques: Activation Assay, Functional Assay, Inhibition, Activity Assay, Phospho-proteomics, Residue, Expressing, Control, Derivative Assay, Fluorescence

Journal: Journal for Immunotherapy of Cancer

Article Title: Preventing surgery-induced natural killer cell suppression and metastases by inhibiting PI3K-gamma signaling in myeloid-derived suppressor cells

doi: 10.1136/jitc-2025-013304

Figure Lengend Snippet: Blockade of PI3Kγ signaling in sx-MDSCs reduces NK cell suppression and metastatic disease in mouse models of surgical stress. ( A ) Ex vivo effect of PI3K inhibitors on pAKT (T308) phosphorylation in splenic MDSCs from C57Bl/6 mice (n=4). ( B ) Effect of preoperative in vivo administration of PI3K inhibitors on pAKT (T308) phosphorylation measured on POD1 in splenic MDSCs (n=4). ( C ) Ex vivo effect of PI3K inhibitors on MDSC-NK %suppression (n=3). ( D ) Effect of preoperative in vivo administration of PI3K-γ inhibitors on sx-MDSC suppressive capacity. ( E ) Adoptive transfer of representative images of lungs is shown. One-way ANOVA with Holm-Sidak or two-way ANOVA with Dunnett’s multiple comparisons tests were performed. ****p<0.0001. ANOVA, analysis of variance; M-MDSC, monocytic-MDSC; MDSC, myeloid-derived suppressor cell; MFI, mean fluorescence intensity; NK, natural killer; sx-MDSC, surgery-induced MDSC, PMN-MDSC, polymorphonuclear-MDSC; POD1, postoperative day 1.

Article Snippet: Sx-MDSCs were isolated using the MDSC Isolation Kit (Miltenyi #130-094-538) from surgically stressed mice on POD1.

Techniques: Ex Vivo, Phospho-proteomics, In Vivo, Adoptive Transfer Assay, Derivative Assay, Fluorescence

Journal: Frontiers in Oncology

Article Title: A novel prognostic signature based on mitochondrial permeability transition-driven necrosis genes for biochemical recurrence prediction in prostate cancer

doi: 10.3389/fonc.2026.1775602

Figure Lengend Snippet: Enrichment analysis and immune-cell infiltration based on risk groups. (A) GSEA analysis showing top 5 pathways between low- and high-risk groups. (B) Heatmap of differences in pathways between low- and high-risk groups. (C) Stacked bar chart showing infiltration proportions of immune cells using ssGSEA. (D) Correlation analysis of FNDC1, S100A8, and TREM2 with differential immune infiltrating cells. (E) Box plot of differences in 7 types of immune cells within the low- and high-risk groups. (F) Correlation analysis between S100A8 and neutrophils. (G) The differences in the scores of dysfunction, exclusion, MSI, and TIDE between low- and high-risk groups. (H) The differences in the expression of immune checkpoint between low- and high-risk groups. Ns, not significant. * indicate p<0.05, ** indicate p<0.01, *** indicate p<0.001, **** indicate p<0.0001.

Article Snippet: The following were the antibody concentrations for western blotting: rabbit polyclonal anti-FNDC1 antibody (1:1000, catalog no. bs-8460R; Bioss, Beijing, China), rabbit polyclonal anti-TREM2 antibody (1:1000, catalog no. HY- P80920 ; MedChemExpress, Monmouth Junction, NJ, USA), and rabbit polyclonal anti-S100A8 antibody (1:1000, catalog no. AF7929; Beyotime, Shanghai, China), and mouse monoclonal anti-GAPDH (1:2000, 60004-1-Ig, Proteintech, China).

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: A novel prognostic signature based on mitochondrial permeability transition-driven necrosis genes for biochemical recurrence prediction in prostate cancer

doi: 10.3389/fonc.2026.1775602

Figure Lengend Snippet: Validation of FNDC1, S100A8, and TREM2 expression. (A) The mRNA expression levels of FNDC1, S100A8, and TREM2 in TCGA-PRAD cohort. (B) The mRNA expression levels of FNDC1, S100A8 and TREM2 were confirmed by q-PCR between paratumor(PT) and prostate cancer tissue. (C, D) The protein expression levels of FNDC1, S100A8 and TREM2 were confirmed by western blot between paratumor(PT) and prostate cancer tissue(T). (E) Immunohistochemistry micrographs revealing FNDC1, S100A8 and TREM2 expression between paratumor(PT) and prostate cancer tissue(T). **P< 0.01; ***P< 0.001.

Article Snippet: The following were the antibody concentrations for western blotting: rabbit polyclonal anti-FNDC1 antibody (1:1000, catalog no. bs-8460R; Bioss, Beijing, China), rabbit polyclonal anti-TREM2 antibody (1:1000, catalog no. HY- P80920 ; MedChemExpress, Monmouth Junction, NJ, USA), and rabbit polyclonal anti-S100A8 antibody (1:1000, catalog no. AF7929; Beyotime, Shanghai, China), and mouse monoclonal anti-GAPDH (1:2000, 60004-1-Ig, Proteintech, China).

Techniques: Biomarker Discovery, Expressing, Western Blot, Immunohistochemistry

Journal: Cell Death and Differentiation

Article Title: JOSD1 inhibits mitochondrial apoptotic signalling to drive acquired chemoresistance in gynaecological cancer by stabilizing MCL1

doi: 10.1038/s41418-019-0339-0

Figure Lengend Snippet: JOSD1 associates with and deubiquitinates myeloid cell leukaemia 1 (MCL1). a Immunoprecipitation was performed using an anti-Flag antibody in ES2 cells expressing empty vector or Flag-JOSD1 and then conducting sliver staining. b Number of unique peptide hits for JOSD1 and MCL1 are shown. c Immunoblotting for V5-MCL1 among proteins pulled down with anti-FLAG M2 affinity gels (left) and for Flag-JOSD1 among proteins pulled down with an anti-V5 antibody (right). d Immunoblotting for the association between HA-JOSD1 and Flag-MCL1 in 293T cells exposed to 0.5 mg/ml carboplatin for 0, 3, 6 and 24 h. e Confocal microscopy of the co-localization of Myc-JOSD1 (green) and MCL1 (red) in ES2 cells. Nuclei were stained with Hoechst 33342 (blue). Scale bars, 60 μm. f Immunoblotting to detect the ubiquitination of MCL1 in 293T cells co-transfected with Myc-JOSD1, HA-Ubiquitin and Flag-MCL1 (wild type or C36A). g Immunoblotting to detect the ubiquitination of MCL1 in A2780 cells expressing either short hairpin RNAs (shRNAs) targeting JOSD1 or nontargeting shRNA. h Immunoblotting to detect the ubiquitination of MCL1 in 293T cells co-transfected with Flag-MCL1, Myc-JOSD1 and HA-Ubiquitin mutant (K0 indicates that all lysines were replaced by arginines, and K48 and K63 indicate that all lysines, except for K48 or K63, were mutated to arginines). Representative results of three biological replicates are shown

Article Snippet: For immunoprecipitation, equal amounts of lysate were incubated with anti-FLAG M2 affinity gels (A2220, Sigma) or anti-V5-tag pAb-agarose (D291-8, MBL, Japan) or with protein A/G magnetic beads (HY-K0202, MCE) and anti-MCL1 antibody overnight at 4 °C.

Techniques: Immunoprecipitation, Expressing, Plasmid Preparation, Staining, Western Blot, Confocal Microscopy, Ubiquitin Proteomics, Transfection, shRNA, Mutagenesis

Journal: Cell Death and Differentiation

Article Title: JOSD1 inhibits mitochondrial apoptotic signalling to drive acquired chemoresistance in gynaecological cancer by stabilizing MCL1

doi: 10.1038/s41418-019-0339-0

Figure Lengend Snippet: JOSD1 stabilizes myeloid cell leukaemia 1 (MCL1) to suppress the mitochondrial apoptosis pathway. a Immunoblotting for MCL1 expression in 293T cells transfected with increasing amounts of JOSD1 or JOSD1C36A. b Immunoblotting for MCL1 expression in 293T cells transfected with JOSD1 or JOSD1C36A and subjected to the cycloheximide (CHX) pulse-chase assay. Quantification of MCL1 expression relative to β-actin expression is shown. c Immunoblotting for MCL1 expression in 293T cells transfected with either JOSD1-targeting or nontargeting siRNAs and subjected to the CHX pulse-chase assay. Quantification of MCL1 relative to β-actin expression is shown. d H-score and representative images of JOSD1 and MCL1 staining in chemoresistant and parental xenografts. Scale bars, 100 μm. e Immunoblotting for apoptosis markers in A2780 and HeLa cells expressing either JOSD1-targeting or nontargeting shRNAs. f Immunoblotting for apoptosis markers in ES2 cells expressing empty vector, JOSD1 or JOSD1C36A and were treated with different concentrations of carboplatin. g In vitro growth of A2780 cells expressing either siRNAs targeting JOSD1 or negative control siRNA and treated with Z-VAD(OMe)-FMK (50 μM) or dimethyl sulfoxide (DMSO). Z-VAD(OMe)-FMK (50 μM) or DMSO was added 2 h before small interfering RNA transfection. Two-tailed Student’s t test; ****p < 0.0001; ns, no significant difference. In b–d, the data are presented as the mean ± SEM; in g, the data are presented as the mean ± SD. Representative results of three biological replicates are shown

Article Snippet: For immunoprecipitation, equal amounts of lysate were incubated with anti-FLAG M2 affinity gels (A2220, Sigma) or anti-V5-tag pAb-agarose (D291-8, MBL, Japan) or with protein A/G magnetic beads (HY-K0202, MCE) and anti-MCL1 antibody overnight at 4 °C.

Techniques: Western Blot, Expressing, Transfection, Pulse Chase, Staining, Plasmid Preparation, In Vitro, Negative Control, Small Interfering RNA, Two Tailed Test

Journal: Cell Death and Differentiation

Article Title: JOSD1 inhibits mitochondrial apoptotic signalling to drive acquired chemoresistance in gynaecological cancer by stabilizing MCL1

doi: 10.1038/s41418-019-0339-0

Figure Lengend Snippet: JOSD1 causes chemoresistance by stabilizing myeloid cell leukaemia 1 (MCL1). a Cell viability of ES2 and 3AO cells expressing empty vector, JOSD1 or JOSD1C36A and treated with different doses of carboplatin (CBP). b Cell viability of ES2 and 3AO cells expressing empty vector, JOSD1 or JOSD1 and MCL1-targeting short hairpin RNAs (shRNAs) and treated with different doses of CBP. c Quantitative analysis of the apoptosis rate of ES2 cells expressing empty vector, JOSD1 or JOSDC36A and treated with different doses of CBP. d–f In vivo growth curves and tumour weights of xenografts generated from ES2 cells expressing an empty vector, JOSD1 or JOSD1 and MCL1-targeting shRNAs and treated with CBP (50 mg/kg, three times a week). g, h H-score and representative images of immunohistochemical staining for JOSD1, MCL1 and cleaved caspase-3 in Fig. 5e. Scale bars, 250 μm. Two-tailed Student’s t test; **p < 0.01, ***p < 0.001 and ****p < 0.0001. In a and b, the data are presented as the mean ± SD, and representative results of three biological replicates are shown. In c, d, f and g, the data are presented as the mean ± SEM (n = 10)

Article Snippet: For immunoprecipitation, equal amounts of lysate were incubated with anti-FLAG M2 affinity gels (A2220, Sigma) or anti-V5-tag pAb-agarose (D291-8, MBL, Japan) or with protein A/G magnetic beads (HY-K0202, MCE) and anti-MCL1 antibody overnight at 4 °C.

Techniques: Expressing, Plasmid Preparation, In Vivo, Generated, Immunohistochemical staining, Staining, Two Tailed Test

Journal: Cell Death and Differentiation

Article Title: JOSD1 inhibits mitochondrial apoptotic signalling to drive acquired chemoresistance in gynaecological cancer by stabilizing MCL1

doi: 10.1038/s41418-019-0339-0

Figure Lengend Snippet: JOSD1 and myeloid cell leukaemia 1 (MCL1) expression correlates with chemoresistance and poor prognosis of ovarian cancer patients. a Representative images of JOSD1 and MCL1 staining in platinum-sensitive and platinum-resistant ovarian cancer samples. Scale bars, 250 μm. b Correlation analysis of JOSD1 and MCL1 expression levels in ovarian cancer samples (n = 150). Pearson correlation coefficients are shown. c, d Correlation between JOSD1 expression (c) or MCL1 expression (d) and overall survival or progression-free survival in ovarian cancer patients (n = 150). Kaplan–Meier survival plots are shown. e Immunoblotting for JOSD1 in the supernatants of chemoresistant cell lines and their parental cell lines. f Correlation between serum JOSD1 levels and intratumour JOSD1 expression in 20 paired serum and tumour tissue samples. Pearson correlation coefficients are shown

Article Snippet: For immunoprecipitation, equal amounts of lysate were incubated with anti-FLAG M2 affinity gels (A2220, Sigma) or anti-V5-tag pAb-agarose (D291-8, MBL, Japan) or with protein A/G magnetic beads (HY-K0202, MCE) and anti-MCL1 antibody overnight at 4 °C.

Techniques: Expressing, Staining, Western Blot

Journal: Cell Death and Differentiation

Article Title: JOSD1 inhibits mitochondrial apoptotic signalling to drive acquired chemoresistance in gynaecological cancer by stabilizing MCL1

doi: 10.1038/s41418-019-0339-0

Figure Lengend Snippet: JOSD1 drives chemoresistance by stabilizing myeloid cell leukaemia 1 (MCL1) and suppressing mitochondrial apoptotic signalling. JOSD1 interacts with and deubiquitinates MCL1. JOSD1 stabilizes MCL1 protein in the cytoplasm, which leads to the inhibition of mitochondrial apoptotic signalling. The cytoplasmic JOSD1 expression level also correlates with the serum JOSD1 expression level

Article Snippet: For immunoprecipitation, equal amounts of lysate were incubated with anti-FLAG M2 affinity gels (A2220, Sigma) or anti-V5-tag pAb-agarose (D291-8, MBL, Japan) or with protein A/G magnetic beads (HY-K0202, MCE) and anti-MCL1 antibody overnight at 4 °C.

Techniques: Inhibition, Expressing

Journal: Acta Neuropathologica Communications

Article Title: Spatial characterisation of TREM2 expression in actively demyelinating multiple sclerosis lesions supports its key roles in lipid metabolic pathways

doi: 10.1186/s40478-026-02241-x

Figure Lengend Snippet: Spatial transcriptomics in hippocampal tissue containing chronic active lesions from two cases (MS13C and MS15A) of secondary progressive MS. a Schematic of spatial transcriptomics workflow using sample integration, clustering and deconvolution functions. b LFB staining and MHC class II immunohistochemistry were used to identify lesioned (yellow outline) areas (2 mm scale bar; left panels). c Cell populations were estimated by deconvolution analysis of spatial gene expression profiles in each cluster. Clusters 0, 4, 11, 14, 15, and 16 contained a higher proportion of glial cells relative to other clusters associated with neuronal cell types in the hippocampus. d Quantitative analysis of spatial distributions of total TREM2 + spots and TREM2 in relation to selected myeloid cell markers ( CD68, CD163, IBA1 ). OPC = Oligodendrocyte progenitor cell, vSMC = Vascular smooth muscle cell, OLG = Oligodendrocyte

Article Snippet: Rabbit anti-human TREM2 (Polyclonal) , Nonspecific , ProSci, USA (13-679) , 1:100.

Techniques: Spatial Transcriptomics, Staining, Immunohistochemistry, Gene Expression

Journal: Acta Neuropathologica Communications

Article Title: Spatial characterisation of TREM2 expression in actively demyelinating multiple sclerosis lesions supports its key roles in lipid metabolic pathways

doi: 10.1186/s40478-026-02241-x

Figure Lengend Snippet: Gene enrichment pathway analysis of module B revealed lesion-associated submodules, with TREM2 enriched exclusively in submodule B. a Submodules of original module B for the two cases MS13C and MS15A allowed the identification of submodules associated with distinct glial cell states. b Heat map of the cellular composition of each submodule for MS13C and MS15A. c Submodule B hub gene network for MS13C revealed TREM2 and proximal genes. d Gene ontology biological process enrichment analysis for each submodule for MS15A and MS13C. e The results of enrichment analysis within the KEGG gene sets database for co-expressed glial submodules for MS13C and MS15A are shown. OPC = Oligodendrocyte progenitor cell, vSMC = Vascular smooth muscle cell, OLG = Oligodendrocyte, GO = Gene ontology

Article Snippet: Rabbit anti-human TREM2 (Polyclonal) , Nonspecific , ProSci, USA (13-679) , 1:100.

Techniques:

Journal: Acta Neuropathologica Communications

Article Title: Spatial characterisation of TREM2 expression in actively demyelinating multiple sclerosis lesions supports its key roles in lipid metabolic pathways

doi: 10.1186/s40478-026-02241-x

Figure Lengend Snippet: Heat map highlighting heterogeneity of TREM2 protein expression in myeloid cells in MS lesions, MS and control NAWM. NAWM and lesion sub-areas were analysed (lesion centres, PLWM and lesion rims). Staining intensity for ORO, CD68+, MHC class II+ and, MBP+ for each tissue are shown. ORO+ cells per ROI (314.74 × 236.06 microns) ranged from 0 to 30 + cells. CD68+ or MHC class II+ cells per ROI (500 × 500 microns) ranged from 0 to 100+ cells. Relative MBP immunohistochemistry was characterised as “light”, “med” and “dark”. The three main cell types analysed were: PVMs (left), parenchymal macrophages (middle) and microglia (right). Images used to calculate the average number of immunolabeled cells per ROI were 312.39 × 312.39 µm. *Tissue lesions provided by Washington University Repository were identified by LFB and MHC class II staining. **MS3E was a case with active MS lesions, which subsequently was identified to have progressive multifocal leukoencephalopathy. This case was not included in subsequent overall analyses of the cohort of MS lesion cases. ***MS15A was a chronic active lesion confirmed by a neuropathologist, however the section provided did not have a clearly identifiable rim. ORO = Oil Red O, MBP = Myelin Basic Protein, NAWM = Normal-appearing white matter, PLWM = Perilesional white matter, n.a = not applicable, ROI = Region of Interest, MHC = MHC class II

Article Snippet: Rabbit anti-human TREM2 (Polyclonal) , Nonspecific , ProSci, USA (13-679) , 1:100.

Techniques: Expressing, Control, Staining, Immunohistochemistry, Immunolabeling

Journal: Acta Neuropathologica Communications

Article Title: Spatial characterisation of TREM2 expression in actively demyelinating multiple sclerosis lesions supports its key roles in lipid metabolic pathways

doi: 10.1186/s40478-026-02241-x

Figure Lengend Snippet: Characterisation of TREM2 expression on microglia and macrophages in active lesions. a Representative Luxol Fast Blue (LFB) and Oil Red O (ORO) images from MS2A highlight the histopathology of an active MS lesion. Lipid-filled cells in the active lesion are highlighted by LFB staining (left; 25 µm scale bar) and ORO staining (middle; yellow dotted box highlights the area of the right image; 10 µm scale bar). Arrows point to LFB staining inside PVMs (possibly indicative of intracellular myelin products—white arrow), ORO + cells in the CNS parenchyma (white star) and lining the vessel (white arrow) (×40 magnification). b Representative immunofluorescence images of macrophages (left), reactive microglia (centre), and PVMs (right) are from active lesions (from MS2A and MS4A) (×40 magnification; 30 µm scale bar). Cropped images are from the highlighted area in the merged overview (yellow square). The area has been split into separate channels: TREM2 (red), CD163 (green) and Iba1 (white). DAPI-stained nuclei (blue) are only seen in the merged overview images. The white arrow marks a triple-labelled TREM2+ CD163+ Iba1+ macrophage. The star marks a rare putative triple-labelled TREM2+ CD163+ Iba1+ microglia. In this panel, other reactive microglia can be seen that are negative for CD163. c TREM2 (red), CD163 (green) and Iba1 (white) triple-labelled macrophages in the choroid plexus (white arrow) from MS15A (20 µm scale bar)

Article Snippet: Rabbit anti-human TREM2 (Polyclonal) , Nonspecific , ProSci, USA (13-679) , 1:100.

Techniques: Expressing, Histopathology, Staining, Immunofluorescence

Journal: Acta Neuropathologica Communications

Article Title: Spatial characterisation of TREM2 expression in actively demyelinating multiple sclerosis lesions supports its key roles in lipid metabolic pathways

doi: 10.1186/s40478-026-02241-x

Figure Lengend Snippet: Perinuclear and plasma membrane distribution of TREM2 respectively in microglia and macrophages in MS lesions. Actively demyelinating tissues were immunolabelled with TREM2 (red), CD163 (green), Iba1 (white), counter-stained with DAPI (blue; merged images), and imaged using a 100× objective with 6-step confocal z-stacks. a Triple labelled PVM–CD163+ TREM2+ Iba1+ (case MS4A). Punctuate discontinuous labelling with the AF1828 antibody in a thin line around the inner surface of the vessel can also be seen, indicative of endothelium (arrow). b TREM2+ CD163-negative microglia (MS2A), c TREM2+ CD163+ putative microglia (MS2A), d TREM2-negative CD163+ putative microglia (MS4A), e CD163+ TREM2+ macrophage (MS2A). f A rare TREM2+ CD163-negative Iba1-negative cell (white arrow) (MS4A). Scale bars = 5 µm

Article Snippet: Rabbit anti-human TREM2 (Polyclonal) , Nonspecific , ProSci, USA (13-679) , 1:100.

Techniques: Clinical Proteomics, Membrane, Staining

Journal: Acta Neuropathologica Communications

Article Title: Spatial characterisation of TREM2 expression in actively demyelinating multiple sclerosis lesions supports its key roles in lipid metabolic pathways

doi: 10.1186/s40478-026-02241-x

Figure Lengend Snippet: Quantification of TREM2+ cells in control NAWM, MS NAWM and MS lesions. a Number of cells (total DAPI nuclei) per mm 2 , b number of microglia/macrophages (Iba1+ and/or CD163+) per mm 2 , c number of TREM2+ microglia/macrophages per mm 2 , and d percentage of TREM2+ microglia/macrophages in distinct tissue areas as indicated. e Proportions of single, double, and triple labelled TREM2+ CD163+ Iba1+ cell populations in distinct tissue areas. The four most abundant cell phenotypes for each area are listed below each chart. Each cell phenotype has been referenced with the same colour across different lesion types as indicate in figure legend. f Immunofluorescence intensity of TREM2 (pink), Iba1 (blue) and CD163 (green) in distinct tissue areas. The size of the individual cell region of interest used to determine intensity ranged from 64 to 295 µm 2 . * P ≤ 0.05; ** P ≤ 0.01; **** P ≤ 0.0001. NAWM = normal-appearing white matter (from MS cases or controls as indicated), LC = lesion centre, PLWM = perilesional white matter, LR = lesion rim

Article Snippet: Rabbit anti-human TREM2 (Polyclonal) , Nonspecific , ProSci, USA (13-679) , 1:100.

Techniques: Control, Immunofluorescence

Journal: Acta Neuropathologica Communications

Article Title: Spatial characterisation of TREM2 expression in actively demyelinating multiple sclerosis lesions supports its key roles in lipid metabolic pathways

doi: 10.1186/s40478-026-02241-x

Figure Lengend Snippet: TREM2 co-expression with lipid metabolism and activation markers. a TREM2 (red), MHC class II (green), and PLIN2 (white) immunolabelling in parenchymal macrophages (top) and perivascular macrophages (bottom) from an active lesion centre (MS2A). b Quantification per mm 2 of number of PLIN2+ cells, PLIN2+ TREM2+ cells and the proportion of all PLIN2+ cells that are TREM2+ in different tissue areas. All TREM2+ cells were PLIN2+. c TREM2 (red), CD68 (green) and TMEM119 (white) indicate TREM2 expression in active microglia in the lesion centre (top) and PLWM (bottom). DAPI nuclei counterstain is shown in the merged channels (100× objective; 10 µm scale bar) (MS4A). NAWM = Normal-appearing white matter, LC = lesion centre, PLWM = Perilesional white matter, LR = lesion rim

Article Snippet: Rabbit anti-human TREM2 (Polyclonal) , Nonspecific , ProSci, USA (13-679) , 1:100.

Techniques: Expressing, Activation Assay

Journal: Acta Neuropathologica Communications

Article Title: Spatial characterisation of TREM2 expression in actively demyelinating multiple sclerosis lesions supports its key roles in lipid metabolic pathways

doi: 10.1186/s40478-026-02241-x

Figure Lengend Snippet: TREM2 and MS4A4A expression in a PML and active MS lesion. a ORO staining of a PML lesion (20× objective; 200 µm scale bar). The insert (left) shows lipid-filled round macrophage infiltrates characteristic of PML (40× objective; 20 µm scale bar). b TREM2 (red), CD163 (green) and Iba1 (white) immunofluorescence with DAPI nuclei counterstain (blue), highlights abundant CD163+ TREM2+ Iba1+ triple labelled macrophages in the PML lesion with TREM2 distributed to the plasma membrane of foamy macrophages (merge panel 30 µm scale bar; right panels 5 µm scale bar). c TREM2 (red), TMEM119 (white), CD163 (green) and DAPI (blue), 20 µm scale bar. d TREM2 (red), MS4A4A (white) and DAPI (blue), 20 µm scale bar. e TREM2 (red), TMEM119 (white), CD163 (green) and DAPI (blue; merged image) indicate TREM2+ microglia in peri-lesional white matter in PML (10 µm scale bar). f TREM2 (red), CD68 (green) and MS4A4A (white) indicate TREM2 and MS4A4A co-expression in PVMs from an active MS lesion centre (MS4A). DAPI nuclei counterstain is shown in the merged channels (100× objective; 10 µm scale bar). Representative images in a–e are of tissue from PML case MS3E. PLWM = Perilesional white matter, PML = Progressive multifocal leukoencephalopathy

Article Snippet: Rabbit anti-human TREM2 (Polyclonal) , Nonspecific , ProSci, USA (13-679) , 1:100.

Techniques: Expressing, Staining, Immunofluorescence, Clinical Proteomics, Membrane