csf1  (R&D Systems)

Journal: Journal of the Peripheral Nervous System

Article Title: Dorsal root ganglia CSF1 + neuronal subtypes have different impact on macrophages and microglia after spared nerve injury

doi: 10.1111/jns.12674

Figure Lengend Snippet: The number of CSF1 + neurons in L3, L4 and L5 DRG varies after SNI. (A, C and E) Representative fluorescent immunohistochemistry images illustrating CSF1 + DRG neurons 7 days after SNI surgery in L3 (A), L4 (C), and L5 (E) DRG (scale bar, 50 μm). (B, D, and F) Bar graphs representing the number of CSF1 + neurons normalized to Control condition. SNI lesion induced a significant increase compared with Sham condition in all DRG (**** p < .0001 for L3 and L4 DRG, and ** p < .01 for L5 DRG). L4 DRG seems more sensitive, since Sham surgery alone also induced a significant increase compared with Control condition (** p < .01). (G) Bar graph representing the comparison between L3, L4, and L5 DRG, showing that the strongest increase of CSF1 + neurons after SNI was in L4 DRG, followed by L3 and L5 (** p < .01 for L3 vs. L5 and **** p < .0001 for L4 vs. L5).

Article Snippet: The blocking step was followed by a 3‐h incubation at room temperature for all IB4 and L5 DRG stainings, or an overnight incubation at 4°C for the rest of L3 and L4 DRG stainings with antibodies against: NF200 (1:500, rabbit, N4142, Sigma, St. Louis, MO, USA), CGRP (1:1000, rabbit, T4032, BMA Biomedicals, Augst, Switzerland), CSF1 (1:300, goat, AF416, R&D Systems, Minneapolis, MN, USA), and biotinylated isolectin B4 (IB4) (1:100, B‐1205, Vector laboratories, Newark, CA, USA) in 5% donkey serum in 1X PBS + 0.1% Triton X.

Techniques: Immunohistochemistry, Control, Comparison

Journal: Journal of the Peripheral Nervous System

Article Title: Dorsal root ganglia CSF1 + neuronal subtypes have different impact on macrophages and microglia after spared nerve injury

doi: 10.1111/jns.12674

Figure Lengend Snippet: Seven days after SNI, CX3CR1 + macrophages cluster around CSF1 + neurons in L3, L4 and L5 DRG. (A–C) Representative fluorescent immunohistochemistry images illustrating CSF1 + neurons (in red), CX3CR1 + macrophages (in green), and merged images, in L3 (A), L4 (B), and L5 (C) DRG. Insets represent high‐magnification images of the selected areas, showing the perineuronal rings of macrophages that surround CSF1 + neurons after the lesion (indicated by white arrows) (scale bar, 50 μm). (D) Bar graph showing mean percentages of CSF1 + neurons surrounded by rings of CX3CR1 + macrophages in all 3 DRG: L4 DRG > L3 DRG > L5 DRG.

Article Snippet: The blocking step was followed by a 3‐h incubation at room temperature for all IB4 and L5 DRG stainings, or an overnight incubation at 4°C for the rest of L3 and L4 DRG stainings with antibodies against: NF200 (1:500, rabbit, N4142, Sigma, St. Louis, MO, USA), CGRP (1:1000, rabbit, T4032, BMA Biomedicals, Augst, Switzerland), CSF1 (1:300, goat, AF416, R&D Systems, Minneapolis, MN, USA), and biotinylated isolectin B4 (IB4) (1:100, B‐1205, Vector laboratories, Newark, CA, USA) in 5% donkey serum in 1X PBS + 0.1% Triton X.

Techniques: Immunohistochemistry

Journal: Journal of the Peripheral Nervous System

Article Title: Dorsal root ganglia CSF1 + neuronal subtypes have different impact on macrophages and microglia after spared nerve injury

doi: 10.1111/jns.12674

Figure Lengend Snippet: SNI increased the incidence of CSF1 + /NF200 + DRG neurons and decreased the number of CSF1 + /CGRP + and CSF1 + /IB4 + DRG neurons. (A, C, and E) Representative fluorescent immunohistochemistry images illustrating CSF1 + /NF200 + , CSF1 + /CGRP + , and CSF1 + /IB4 + 7 days after SNI in L3 (A), L4 (C), and L5 (E) DRG (scale bar, 50 μm). (B, D, and F) Bar graphs representing the percentages of CSF1 + /NF200 + , CSF1 + /CGRP + , and CSF1 + /IB4 + 7 days after SNI in L3 (B), L4 (D), and L5 (F) DRG. Although there is an increasing trend for CSF1 + /NF200 + and a decreasing trend for CSF1 + /CGRP + and CSF1 + /IB4 + in all DRG, only in L3 DRG there is a significant decrease for CSF1 + /IB4 + neurons. (G) Bar graph showing mean percentages of CSF1 + /NF200 + , CSF1 + /CGRP + , and CSF1 + /IB4 + neurons in all 3 DRG 7 days after SNI. There were no differences between L3, L4, and L5 DRG for each neuronal types.

Article Snippet: The blocking step was followed by a 3‐h incubation at room temperature for all IB4 and L5 DRG stainings, or an overnight incubation at 4°C for the rest of L3 and L4 DRG stainings with antibodies against: NF200 (1:500, rabbit, N4142, Sigma, St. Louis, MO, USA), CGRP (1:1000, rabbit, T4032, BMA Biomedicals, Augst, Switzerland), CSF1 (1:300, goat, AF416, R&D Systems, Minneapolis, MN, USA), and biotinylated isolectin B4 (IB4) (1:100, B‐1205, Vector laboratories, Newark, CA, USA) in 5% donkey serum in 1X PBS + 0.1% Triton X.

Techniques: Immunohistochemistry

Journal: bioRxiv

Article Title: Plasminogen activator inhibitors orchestrate the immunosuppressive tumor microenvironment in pancreatic cancer

doi: 10.1101/2025.06.11.659098

Figure Lengend Snippet: a , Representative H&E and Fibrin(ogen) IHC staining of tumor sections from orthotopically transplanted control, Serpinb2 KO, and Serpine1 KO mice. Scale bar: 50 µm. b , Quantification of Fibrin(ogen) IHC as a percentage of tissue area (control n=4, Serpinb2 KO n=4, Serpine1 KO n=4). c , Pseudocolored immunohistochemistry of tumors from control mice, isolated two weeks post-transplantation. Sections were stained for CK19, CD11b, LY6G, F4/80, Fibrin(ogen), PAI1, and PAI2. Representative images are shown. Scale bar: 200 µm for the main image and 50 µm for insets. d , Comparison of distances between CD11b+F4/80+ cells co-localizing with Fibrin(ogen) (CD11b+F4/80+/Fibrin(ogen)), and CK19+, PAI1+CK19+, and PAI2+CK19+ cells in control tumors (n=6). Only the tumor center area, as visualized in figure S9 , panel ( a ), was used for the analysis. e , Density plot showing the spatial distribution of CK19+ (grey), PAI2+CK19+ (red) and PAI1+CK19+ (purple) cells relative to CD11b+F4/80+/Fibrin(ogen) immune cells. The x-axis shows the distance (μm) from CD11b+F4/80+/Fibrin(ogen) cells and the y-axis the distribution density. Only the tumor center area, as visualized in figure S9 , panel ( a ), was used for the analysis. f , Schematic of the experimental design. Mice were pre-treated with IgG+PBS-liposomes or anti CSF1+clodronate and orthotopically transplanted with control, Serpinb2 KO, or Serpine1 KO KPC cells. Mice were treated twice weekly for two weeks. Tumors were harvested for phenotypic analysis. g , Tumor weight comparison between IgG+PBS-liposome control (n=4), Serpinb2 KO (n=4), and Serpine1 KO (n=4) tumors, and anti CSF1+clodronate-liposome control (n=4), Serpinb2 KO (n=3), and Serpine1 KO (n=4) tumors. h , Flow cytometry analysis of tumors isolated from IgG+PBS-liposome control, and anti CSF1+clodronate-liposome control, Serpinb2 , and Serpine1 KO mice two weeks after orthotopic transplantation. The absolute numbers of macrophages are shown (IgG+PBS-liposome control n=4, anti CSF1+clodronate-liposome control n=4, anti CSF1+clodronate-liposome Serpinb2 KO n=3, anti CSF1+clodronate-liposome Serpine1 KO n=3). Statistical significance in b , g and h was determined using a two-tailed, unpaired Student’s t-test. P value in d was calculated with a two-tailed, paired t-test KO: knock-out, IHC: immunohistochemistry

Article Snippet: In the macrophage depletion group, mice were treated with 1 mg of InVivoMAb anti-mouse CSF1 (anti-CSF1, #BE0204, Bioxcell) on day 1 and 200 μl of clodronate liposomes (SKU# CLD-8914, Encapsula) on day 2.

Techniques: Immunohistochemistry, Control, Isolation, Transplantation Assay, Staining, Comparison, Liposomes, Flow Cytometry, Two Tailed Test, Knock-Out

Journal: Cell Reports Medicine

Article Title: CV1-secreting sCAR-T cells potentiate the abscopal effect of microwave ablation in heterogeneous tumors

doi: 10.1016/j.xcrm.2025.101965

Figure Lengend Snippet: The enhanced abscopal effect induced by MAsCART treatment relies mainly on macrophages, rather than on innate T cells (A) Hepa1-6-chGPC3 or SK-HEP-1-GPC3 cells were engrafted into the right (local) and Hepa1-6 or SK-HEP-1 cells were inoculated into the left (distant) flank of C57BL/6J or NOG mice, respectively. Anti-CD8a or CSF1 antibody was used in combination treatment accordingly. Tumor-bearing C57BL/6J mice received lymphodepletion (LD) on the day before treatment. (B–E) Evaluation of the abscopal effect in T cell-deficient mice ( n = 5 of each, one experiment). Flow cytometry analysis (B) to detect the T cell depletion in the spleen with anti-CD8a antibody in C57BL/6J mice (C). Distant tumor (Hepa1-6) growth followed by MWA and MAmsCART with/without anti-CD8a antibody treatment in C57BL/6J mice (D). Distant tumor (SK-HEP-1) growth followed by MWA and MAsCART treatment in NOG mice (E). (F–J) Evaluation of the abscopal effect in macrophage-deficient mice. Tumor growth of distant non-ablated tumor (Hepa1-6) in C57BL/6J mice by MAmsCART with/without anti-CSF1 antibody, n = 5, one experiment (F). Tumor-infiltrating macrophages in MAmsCART ± anti-CSF1 antibody groups were detected via flow cytometry, n = 5, one experiment (G and H). Distant tumor (SK-HEP-1) growth followed by MWA and MAsCART with/without anti-CSF1 antibody in NOG mice, n = 5, one experiment (I). Tumor-infiltrating macrophages in distant non-ablated tumor (SK-HEP-1) in NOG mice with MAsCART ± anti-CSF1 treatment by flow cytometry on day 6, n = 5, one experiment (J). (K and L) t-SNE plot showing the subtypes of inflammatory macrophages (K) and their percentages in sequenced cells (L) derived from the MACART and MAsCART group in Figure 3 A, n = 3, one experiment. Error bars are means ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 by two-way ANOVA with Tukey’s multiple comparisons test for (D), (E), (F), and (I) or t test for (C), (H), (J), and (L).

Article Snippet: anti-CSF1 antibody (5A1) , BioXCell , Cat#BE0204; RRID: AB_10950309.

Techniques: Flow Cytometry, Derivative Assay

Journal: Cell Reports Medicine

Article Title: CV1-secreting sCAR-T cells potentiate the abscopal effect of microwave ablation in heterogeneous tumors

doi: 10.1016/j.xcrm.2025.101965

Figure Lengend Snippet: Released chemokines post-MWA by residual tumor are critical for the recruitment of macrophages that elicited the abscopal effect (A) Cytokine array from water-bathed (48°C, 10 min) and control SK-HEP-1-GPC3 human liver cancer cells on days 1, 3, and 6 by bulk RNA sequencing, n = 3, one experiment. (B) CSF1, CCL26, and CXCL8 secretion in water-bathed supernatant of SK-HEP-1-GPC3 after 48 h by ELISA, n = 3 independent experiments. (C) Normalized migration ability of mouse macrophages cultured with conditioned supernatant of water-bathed or control SK-HEP-1-GPC3 cells, n = 3 independent experiments. (D–G) Relative C sf1 mRNA level by qPCR in control and Csf1 knockout SK-HEP-1-GPC3 cells, n = 3 independent experiments (D). Distant tumor (SK-HEP-1) growth followed by local MWA and local SK-HEP-1-GPC3 Csf1 knockout ± MAsCART in NOG mice, n = 5, one experiment (E and F). Distant tumor-infiltrating macrophages in different groups following local MWA and local SK-HEP-1-GPC3 Csf1 knockout ± MAsCART in NOG mice on day 6, n = 5, one experiment (G). Error bars are means ± SD (B, C, D, F, and G). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 by t test for (C) and (D), two-way (F), or one-way (G) ANOVA with Tukey’s multiple comparisons test. See also Figure S4 .

Article Snippet: anti-CSF1 antibody (5A1) , BioXCell , Cat#BE0204; RRID: AB_10950309.

Techniques: Control, RNA Sequencing Assay, Enzyme-linked Immunosorbent Assay, Migration, Cell Culture, Knock-Out

Journal: Cell Reports Medicine

Article Title: CV1-secreting sCAR-T cells potentiate the abscopal effect of microwave ablation in heterogeneous tumors

doi: 10.1016/j.xcrm.2025.101965

Figure Lengend Snippet:

Article Snippet: anti-CSF1 antibody (5A1) , BioXCell , Cat#BE0204; RRID: AB_10950309.

Techniques: Recombinant, Protease Inhibitor, Selection, Cell Isolation, Enzyme-linked Immunosorbent Assay, RNA Extraction, Software