Journal: bioRxiv

Article Title: Therapy-induced senescent-like cancer cells drive macrophage-mediated immunosuppression in cholangiocarcinoma

doi: 10.64898/2026.06.24.734341

Figure Lengend Snippet: (A) Proteome Profiler Human XL cytokine array (105 cytokines) of conditioned medium from proliferating, doxorubicin-induced (Dox), and Gem/Cis-induced senescent-like RBE cells. (B) Gdf15 mRNA in proliferating versus senescent-like murine CCA (SB1) cells. (C) Gdf15 mRNA in control vs AP-treated SB-p16-ATTAC tumors. (D) Schematic of in vitro studies. (E-F) Arg1 and iNOS mRNA in bone-marrow-derived macrophage (BMDM) incubated for 72 h with conditioned medium from vehicle- or alisertib-treated SB1 cells expressing shCtrl or shGdf15 . (G) Bulk RNA-seq heatmap (row Z-scores) of naïve bone marrow-derived macrophages (BMDM) cultured 72 h with conditioned media (CM) from SB-p16-shCtrl or SB- p16-Gdf15 OE cells. (H) ARG1 mRNA in human monocyte-derived macrophages ± recombinant human GDF-15 (Qkine Qk017; TGF-β1-free), qRT-PCR relative to vehicle; mean ± SEM, n = 6. (I) Division index (CFSE dilution, normalized to T-cells-only) of anti-CD3/CD28-activated CD8⁺ T cells co-cultured with BMDMs incubated for 72 h with conditioned medium from vehicle- or alisertib-treated SB1 cells expressing shCtrl or shGdf15 . Data are mean ± SEM; *, P<0.05; **, P<0.01; ****, P<0.0001; ns, not significant.

Article Snippet: Human recombinant GDF-15 was from Qkine (Qk017) and murine recombinant GDF-15 from R&D Systems (8944-GD/CF).

Techniques: Control, In Vitro, Derivative Assay, Incubation, Expressing, RNA Sequencing, Cell Culture, Recombinant, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Therapy-induced senescent-like cancer cells drive macrophage-mediated immunosuppression in cholangiocarcinoma

doi: 10.64898/2026.06.24.734341

Figure Lengend Snippet: (A) Macrophages express TGFBR2, the candidate non-canonical GDF-15 receptor, in murine and human CCA single-cell RNA-seq. (B) α-TGFBR2 western blot (WB) of streptavidin pull-down (top) and input (middle) from naïve BMDMs treated with biotin-supplemented conditioned media (CM) from CCA cells expressing GDF-15-BioID2-HA or SEC61B-BioID2-HA (ER control); α-HA blot of CM (bottom) confirms bait secretion. (C) GDF-15 induces macrophage phospho-STAT6 but not phospho-SMAD2/3, reflecting non-canonical TGFBR2 output. (D) WB of BMDMs stimulated with control or GDF-15-overexpressing (GDF-15 OE ) CM for 0 or 1 h: p-STAT6, STAT6, p-SMAD3, SMAD3. (E) Arg1 mRNA in BMDMs transfected with siCtrl or siTgfbr2 and stimulated with control or rGDF-15. n = 6. Data are mean ± SEM; *, P<0.05; **, P<0.01; ****, P<0.0001; ns, not significant.

Article Snippet: Human recombinant GDF-15 was from Qkine (Qk017) and murine recombinant GDF-15 from R&D Systems (8944-GD/CF).

Techniques: Single Cell, RNA Sequencing, Western Blot, Expressing, Control, Transfection

Journal: bioRxiv

Article Title: Therapy-induced senescent-like cancer cells drive macrophage-mediated immunosuppression in cholangiocarcinoma

doi: 10.64898/2026.06.24.734341

Figure Lengend Snippet: (A) Strategy for p16-promoter-restricted Gdf15 silencing (shRNA) and Gdf15 overexpression. (B) qPCR validation of Gdf15 knockdown and overexpression in senescent-like SB1 cells. (C-D) Subcutaneous model and tumor-burden quantification. (E) GeoMx spatial profiling of GDF-15 expression and macrophage infiltration in human CCA. Region-of-interest (ROI)-level analysis plotting GDF-15 expression against macrophage score; each point is one ROI, with the linear fit and 95% confidence band shown. Spearman ρ = 0.63, P = 0.0067. Data are mean ± SEM; **, P<0.01; ****, P<0.0001.

Article Snippet: Human recombinant GDF-15 was from Qkine (Qk017) and murine recombinant GDF-15 from R&D Systems (8944-GD/CF).

Techniques: shRNA, Over Expression, Biomarker Discovery, Knockdown, Expressing

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Activated M2 Macrophages Contribute to the Pathogenesis of IgG4‐Related Disease via Toll‐like Receptor 7/Interleukin‐33 Signaling

doi: 10.1002/art.41052

Figure Lengend Snippet: Identification of TLR ‐7–expressing cells in SMG s from patients with IgG4‐ RD and in normal secondary lymphoid organs from patients with oral squamous cell carcinoma. A , Staining of serial sections of normal tonsil, normal lymph node, and IgG4‐ RD SMG s with H&E, and for TLR ‐7, CD 68 as a marker of both M1 and M2 macrophages, CD 163 as a marker of M2 macrophages, CD 11c as a marker of myeloid dendritic cells, and CD 123 as a marker of plasmacytoid dendritic cells. Outlined areas indicate GC s. Mayer's hematoxylin (blue) counterstained; bars = 100 μm. B , Double immunostaining for TLR ‐7 (red), CD 123 (green), and CD 163 (green) in normal tonsil and SMG s from representative patients with IgG4‐ RD . DAPI (blue) counterstained; bars = 50 μm. See Figure for definitions.

Article Snippet: Cells (5 × 10 6 cells/ml) were cultured using a CellXVivo Human M2 Macrophage Differentiation Kit (R&D Systems) for 6 days in a humidified chamber with 5% CO 2 at 37°C.

Techniques: Expressing, Staining, Marker, Double Immunostaining

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Activated M2 Macrophages Contribute to the Pathogenesis of IgG4‐Related Disease via Toll‐like Receptor 7/Interleukin‐33 Signaling

doi: 10.1002/art.41052

Figure Lengend Snippet: Expression of interleukin‐33 ( IL ‐33) and candidate TLR s in SG s from patients with IgG4‐ RD . A , Expression levels of mRNA for IL ‐33 in SG s from healthy controls (n = 10), patients with chronic sialadenitis (n = 10), patients with SS (n = 15), and patients with IgG4‐ RD (n = 15). B , Distribution of IL ‐33 in SG s from a representative healthy control, patient with chronic sialadenitis, patient with SS , and patient with IgG4‐ RD . Mayer's hematoxylin (blue) counterstained; bars = 100 μm. C , Correlation between expression levels of IL ‐33 mRNA and candidate TLR s in SG s from patients with IgG4‐ RD (n = 15), as determined by Spearman's rank correlation test. D , Schematic illustration of the extraction of CD 163+ M2 macrophages stimulated with TLR ‐7 agonist R848. Cells were cultivated as described in Patients and Methods. PBMC = peripheral blood mononuclear cell. E , Production of IL ‐33 by CD 163+ M2 macrophages stimulated with R848, as determined by enzyme‐linked immunosorbent assay. IL ‐33 levels increased in a concentration‐dependent manner. In A and E , bars show the mean ± SD . * = P < 0.05; ** = P < 0.01 by Kruskal‐Wallis test. See Figure for other definitions.

Article Snippet: Cells (5 × 10 6 cells/ml) were cultured using a CellXVivo Human M2 Macrophage Differentiation Kit (R&D Systems) for 6 days in a humidified chamber with 5% CO 2 at 37°C.

Techniques: Expressing, Control, Extraction, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Activated M2 Macrophages Contribute to the Pathogenesis of IgG4‐Related Disease via Toll‐like Receptor 7/Interleukin‐33 Signaling

doi: 10.1002/art.41052

Figure Lengend Snippet: Schematic model of the role of Toll‐like receptor 7 ( TLR ‐7)–positive M2 macrophages in the initiation of Ig4‐related disease. TLR ‐7 expressed on M2 macrophages recognize some RNA viruses or self RNA from apoptotic cells. Activated M2 macrophages secrete interleukin‐33 ( IL ‐33) and promote production of Th2 cytokines that lead to IgG4 class‐switching and fibrosis. TGF β = transforming growth factor β.

Article Snippet: Cells (5 × 10 6 cells/ml) were cultured using a CellXVivo Human M2 Macrophage Differentiation Kit (R&D Systems) for 6 days in a humidified chamber with 5% CO 2 at 37°C.

Techniques: