Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Decorin facilitates T cell-mediated antitumor immunity and augments the efficacy of anti-PD1 immunotherapy

doi: 10.1007/s00262-025-04290-y

Figure Lengend Snippet: DCN increases the efficacy of anti-PD1 treatment. A - C C57BL/6 mice were implanted with DCN-overexpressed or control KP141 tumor cells with treatment of IgG isotype control or anti-PD1 monoclonal antibody ( n = 8 from 4 mice per group). The experimental schematic diagram ( A ), tumor volume ( B ), and tumor weight ( C ). D Immunofluorescent staining of CD8 and DAPI in subcutaneous KP141 tumors with or without DCN overexpression. Quantification of CD8 + cells per field is shown on the right ( n = 10). E The relative expression levels of Dcn in responders and non-responders to ICB therapy were analyzed across multiple mouse tumor models using the analytical platform at tismo.cistrome.org. F Kaplan–Meier survival analysis of OS in patients with ICB therapy. Two-sided log-rank test. G Kaplan–Meier survival curves for OS in patients with anti-PD1 therapy. Data from kmplot.com. Two-sided log-rank test

Article Snippet: In vivo anti-PD1 treatment: anti-mouse PD1 (BE0273, BioXcell) and anti-rat IgG2a isotype control (BE0089, BioXcell).

Techniques: Control, Staining, Over Expression, Expressing

Journal: The Journal of Experimental Medicine

Article Title: The SIRPα–CD47 immune checkpoint in NK cells

doi: 10.1084/jem.20200839

Figure Lengend Snippet: Cd47 overexpression protects MHC-deficient mouse iPSCs from killing by stimulated NK cells or macrophages in vivo. A 1:1 mixture of CFSE-labeled WT and either B2m − / − Ciita − / − or B2m − / − Ciita − / − Cd47 tg miPSCs was injected into the peritoneum of mice on a syngeneic C57BL/6 background. After 48 h, the ratio of recovered CFSE-positive miPSCs was determined (mean ± SD, triplicates in four animals per group). (A) Recipient WT C57BL/6 mice did not receive additional treatment. (B) A tg CD11b-DT receptor mouse on C57BL/6 background was used to selectively deplete macrophages. (C) The peritoneal cell populations in macrophage-depleted tg CD11b-DT receptor mice were restored by peritoneal cell transfer from WT C57BL/6 mice. (D) Macrophages were pharmacologically depleted in WT C57BL/6 mice using clodronate. (E) In macrophage-depleted mice, peritoneal NK cells were stimulated by peritoneal injections of mouse IL-2. (F) In macrophage-depleted mice, peritoneal NK cells were stimulated by injections of mouse IL-15. (G) In C57BL/6 mice, NK cells were depleted with an anti-NK1.1 depleting antibody. (H) In WT C57BL/6 mice, both macrophages and NK cells were depleted. (I) A 1:1 mixture of CFSE-labeled B2m − / − Ciita − / − and B2m − / − Ciita − / − Cd47 tg miPSCs was injected into WT C57BL/6 mice. wt, wild-type.

Article Snippet: In some cases, cells were stimulated with mouse IL-2 (1 ng/ml; PeproTech) or stimulator cells were pretreated with anti-mouse Cd47 blocking antibody (clone MIAP301, rat IgG2a,κ; BioXCell) and effector cells with mouse FcR block (catalog no. 130–092-575, concentration 1:5; Miltenyi) for 120 min at 37°C.

Techniques: Over Expression, In Vivo, Labeling, Injection

Journal: The Journal of Experimental Medicine

Article Title: The SIRPα–CD47 immune checkpoint in NK cells

doi: 10.1084/jem.20200839

Figure Lengend Snippet: Cd47 protects MHC-deficient miECs and CD47 protects HLA-deficient hiECs from killing by NK cells and macrophages in vitro. (A–D) WT, B2m − / − Ciita − / − , and B2m − / − Ciita − / − Cd47 tg miECs were challenged with syngeneic C57BL/6 NK cells (A), allogeneic BALB/c NK cells (B), syngeneic C57BL/6 macrophages (C), or allogeneic BALB/c macrophages (D). Where indicated, NK cells were stimulated with mouse IL-2 or IL-15. Graphs show mean ± SD and three independent replicates per group and time point; three different E:T ratios are shown. (E) B2m − / − Ciita − / − miECs were incubated with syngeneic C57BL/6 macrophages treated with an activating anti-Pirb antibody (mean ± SD, three independent replicates per group and time point, and three different E:T ratios are shown). (F and G) WT, B2M − / − CIITA − / − , and B2M − / − CIITA − / − CD47 tg hiECs were challenged with allogeneic human primary NK cells (F) or allogeneic human macrophages (G). Where indicated, NK cells were stimulated with IL-2 or IL-15. Where indicated, an agonist anti-LILRB1 antibody (clone GHI/75) was added to the assay. A specific blocking antibody against CD47 (clone B6.H12) or a blocking peptide against SIRPα was used in some assays. Graphs show mean ± SD and three independent replicates per group and time point; three different E:T ratios are shown. wt, wild-type.

Article Snippet: In some cases, cells were stimulated with mouse IL-2 (1 ng/ml; PeproTech) or stimulator cells were pretreated with anti-mouse Cd47 blocking antibody (clone MIAP301, rat IgG2a,κ; BioXCell) and effector cells with mouse FcR block (catalog no. 130–092-575, concentration 1:5; Miltenyi) for 120 min at 37°C.

Techniques: In Vitro, Incubation, Blocking Assay

Journal: The Journal of Experimental Medicine

Article Title: The SIRPα–CD47 immune checkpoint in NK cells

doi: 10.1084/jem.20200839

Figure Lengend Snippet: The protective effect of Cd47 overexpression against NK cell and macrophage killing is mediated through Sirpα. (A) Sirpα expression on C57BL/6 macrophages and time course of Sirpα expression on C57BL/6 NK cells stimulated with mouse IL-2 (mean ± SD, four independent experiments per group, ANOVA). (B) Cd47 binding to C57BL/6 macrophages and time course of Cd47 binding to C57BL/6 NK cells stimulated with mouse IL-2 (mean ± SD, four independent experiments per group, ANOVA). (C) Sirpα expression on naive C57BL/6 NK cells and 48 h after in vivo stimulation with i.p. mouse IL-2 (mean ± SD, six independent experiments per group, Student’s t test). (D–I) A 1:1 mixture of CFSE-labeled WT and B2m − / − Ciita − / − Cd47 tg miPSCs was injected into the peritoneum of syngeneic C57BL/6 mice and after 48 h, and the ratio of recovered CFSE-positive miPSCs was determined (mean ± SD, triplicates in four animals per group). Animals after NK cell depletion received an anti-Cd47 blocking antibody (clone BE0270) with the miPSC injection (D). Animals after macrophage depletion received an anti-Cd47 blocking antibody with the miPSC injection (E) and additionally mouse IL-2 to activate NK cells in vivo (F). Animals after NK cell depletion received an anti-Sirpα blocking antibody (clone P84; G). Animals after macrophage depletion received an anti-Sirpα blocking antibody (H) and additionally mouse IL-2 to activate NK cells in vivo (I). (J) Sirpα expression on Sirpa − / − macrophages and time course of Sirpα expression on Sirpa − / − NK cells stimulated with mouse IL-2 (mean ± SD, four independent experiments per group, ANOVA). (K) Cd47 binding to Sirpa − / − macrophages and time course of Cd47 binding to Sirpa − / − NK cells stimulated with mouse IL-2 (mean ± SD, four independent experiments per group, ANOVA). (L–O) In some Sirpa − / − mice, NK cells were depleted (L). In other Sirpa − / − mice, macrophages were depleted, and NK cells were stimulated with mouse IL-2 (M); some animals in addition received a blocking antibody for Sirpα (N) or Cd47 (O). Graphs show mean ± SD and triplicates in four animals per group. (P) WT, B2m − / − Ciita − / − , and B2m − / − Ciita − / − Cd47 tg miECs were challenged with Sirpa − / − macrophages (mean ± SD, three independent replicates per group and time point, and three different E:T ratios). (Q) B2m − / − Ciita − / − Cd47 tg miECs were challenged with mouse IL-2–stimulated C57BL/6 NK cells or Sirpa − / − NK cells. In some groups, an anti-Cd47 or anti-Sirpα blocking antibody was used (mean ± SD, three independent replicates per group and time point, and three different E:T ratios). (R) IFNγ ELISpot assays with B2m − / − Ciita − / − Cd47 tg miEC target cells and either mouse IL-2–stimulated C57BL/6 or Sirpa − / − NK cells. Yac-1 served as controls (boxes show 25th to 75th percentile with median, and whiskers show minimum to maximum; six independent samples per group, Student’s t test). wt, wild-type.

Article Snippet: In some cases, cells were stimulated with mouse IL-2 (1 ng/ml; PeproTech) or stimulator cells were pretreated with anti-mouse Cd47 blocking antibody (clone MIAP301, rat IgG2a,κ; BioXCell) and effector cells with mouse FcR block (catalog no. 130–092-575, concentration 1:5; Miltenyi) for 120 min at 37°C.

Techniques: Over Expression, Expressing, Binding Assay, In Vivo, Labeling, Injection, Blocking Assay, Enzyme-linked Immunospot

Journal: The Journal of Experimental Medicine

Article Title: The SIRPα–CD47 immune checkpoint in NK cells

doi: 10.1084/jem.20200839

Figure Lengend Snippet: Expression of SIRPα and CD47 binding of mouse and primary human NK cells. (A and B) The kinetics of Sirpα expression (A) and Cd47 binding (B) of C57BL/6 NK cells in the presence of mouse IL-2 was assessed by flow cytometry (representative histograms are shown of four independent experiments). (C and D) The kinetics of Sirpα expression (C) and Cd47 binding (D) of Sirpa − / − NK cells in the presence of mouse IL-2 was assessed by flow cytometry (representative histograms are shown of four independent experiments). (E and F) The kinetics of SIRPα expression (E) and CD47 binding (F) of human primary NK cells in the presence of IL-2 was assessed by flow cytometry (representative histograms are shown of four independent experiments). (G) The expression of CD47 on five B2M − / − CIITA − / − CD47 tg hiEC clones was assessed by flow cytometry (representative histograms of four independent experiments are shown). wt, wild-type.

Article Snippet: In some cases, cells were stimulated with mouse IL-2 (1 ng/ml; PeproTech) or stimulator cells were pretreated with anti-mouse Cd47 blocking antibody (clone MIAP301, rat IgG2a,κ; BioXCell) and effector cells with mouse FcR block (catalog no. 130–092-575, concentration 1:5; Miltenyi) for 120 min at 37°C.

Techniques: Expressing, Binding Assay, Flow Cytometry, Clone Assay

Journal: The Journal of Experimental Medicine

Article Title: The SIRPα–CD47 immune checkpoint in NK cells

doi: 10.1084/jem.20200839

Figure Lengend Snippet: CD47 is species specific, with no cross-reactivity between mouse and human. (A and B) IFN-γ ELISpot assays were performed with B2m − / − Ciita − / − and B2m − / − Ciita − / − Cd47 tg miPSCs as target cells and syngeneic C57BL/6 (A) or xenogeneic human primary NK cells as effector cells (B). Cd47 antibody blockade (clone BE0270) was used in some groups, and Yac-1 was used as a control (boxes show 25th to 75th percentile with median, and whiskers show minimum to maximum; 12 independent experiments per miPSC group and 6 for Yac-1 groups, ANOVA with Bonferroni’s post hoc test). (C and D) IFN-γ ELISpot assays were performed with B2M − / − CIITA − / − and B2M − / − CIITA − / − CD47 tg hiPSCs as target cells and allogeneic human primary NK cells (C) or xenogeneic C57BL/6 NK cells as effector cells (D). CD47 antibody blockade (clone B6.H12) was used in some groups, and K562 was used as a control (boxes show 25th to 75th percentile with median, and whiskers show minimum to maximum, 12 independent experiments per hiPSC group and 6 for K562 groups, ANOVA with Bonferroni’s post hoc test). (E–H) Fluc + B2M − / − CIITA − / − and B2M − / − CIITA − / − CD47 tg hiPSCs were incubated with allogeneic human macrophages (E and F) or xenogeneic C57BL/6 macrophages (G and H), and the BLI signal was quantified (boxes show 25th to 75th percentile with median, and whiskers show minimum to maximum; n = 16 [control], 20 [macrophages], and 9 [Triton X-100] independent samples, ANOVA with Bonferroni’s post hoc test). CD47 antibody blockade was used in some groups. (I–L) Fluc + B2m − / − Ciita − / − and B2m − / − Ciita − / − Cd47 tg miPSCs were incubated with syngeneic C57BL/6 macrophages (I and J) or xenogeneic human macrophages (K and L), and the BLI signal was quantified (boxes show 25th to 75th percentile with median, and whiskers show minimum to maximum; n = 16 [control], 20 [macrophages], and 9 [Triton X-100] independent samples, ANOVA with Bonferroni’s post hoc test). Cd47 antibody blockade was used in some groups.

Article Snippet: In some cases, cells were stimulated with mouse IL-2 (1 ng/ml; PeproTech) or stimulator cells were pretreated with anti-mouse Cd47 blocking antibody (clone MIAP301, rat IgG2a,κ; BioXCell) and effector cells with mouse FcR block (catalog no. 130–092-575, concentration 1:5; Miltenyi) for 120 min at 37°C.

Techniques: Enzyme-linked Immunospot, Control, Incubation

Journal: The Journal of Experimental Medicine

Article Title: The SIRPα–CD47 immune checkpoint in NK cells

doi: 10.1084/jem.20200839

Figure Lengend Snippet: Threshold of CD47 expression for NK cell and macrophage inhibition. (A) SIRPα expression on macrophages and time course of SIRPα expression on highly selected CD3 − CD7 + CD56 + primary NK cells stimulated with IL-2 (mean ± SD, four independent experiments per group, ANOVA). (B) CD47 binding to macrophages and time course of CD47 binding to highly selected CD3 − CD7 + CD56 + primary NK cells stimulated with IL-2 (mean ± SD, four independent experiments per group, ANOVA). (C and D) From the pool of B2M − / − CIITA − / − CD47 tg hiECs, five clones with different levels of CD47 overexpression were selected. The CD47 expression on transduced cells was always higher than the basal expression level on WT hiECs but below or above the mean of the pool. CD47 was quantified by RT-PCR (C; mean ± SD, three independent experiments per group) and fluorescence (D; mean ± SD, four independent experiments per group). (E and F) The five B2M − / − CIITA − / − CD47 tg hiEC clones were challenged with IL-2–activated human primary NK cells (E) or human macrophages (F). Graphs show mean ± SD and three independent replicates per group and time point; three different E:T ratios are shown. (G) A 1:1 mixture of CFSE-labeled WT and one of the B2M − / − CIITA − / − CD47 tg hiEC clones was injected into the peritoneum of immunodeficient NSG mice. Additionally, either IL-2–activated primary NK cells or macrophages were coinjected. After 48 h, the ratio of recovered CFSE-positive hiECs was determined (mean ± SD, triplicates in four animals per group). wt, wild-type.

Article Snippet: In some cases, cells were stimulated with mouse IL-2 (1 ng/ml; PeproTech) or stimulator cells were pretreated with anti-mouse Cd47 blocking antibody (clone MIAP301, rat IgG2a,κ; BioXCell) and effector cells with mouse FcR block (catalog no. 130–092-575, concentration 1:5; Miltenyi) for 120 min at 37°C.

Techniques: Expressing, Inhibition, Binding Assay, Clone Assay, Over Expression, Reverse Transcription Polymerase Chain Reaction, Fluorescence, Labeling, Injection

Journal: The Journal of Experimental Medicine

Article Title: The SIRPα–CD47 immune checkpoint in NK cells

doi: 10.1084/jem.20200839

Figure Lengend Snippet: Expression of SIRPα and CD47 binding of human NK cell lines. (A and B) The kinetics of SIRPα expression (A) and CD47 binding (B) of human NK cell lines after 24 h of IL-2 stimulation was assessed by flow cytometry (representative histograms are shown of four independent experiments, bar graphs show mean ± SD, four independent experiments per group, ANOVA).

Article Snippet: In some cases, cells were stimulated with mouse IL-2 (1 ng/ml; PeproTech) or stimulator cells were pretreated with anti-mouse Cd47 blocking antibody (clone MIAP301, rat IgG2a,κ; BioXCell) and effector cells with mouse FcR block (catalog no. 130–092-575, concentration 1:5; Miltenyi) for 120 min at 37°C.

Techniques: Expressing, Binding Assay, Flow Cytometry

Journal: The Journal of Experimental Medicine

Article Title: The SIRPα–CD47 immune checkpoint in NK cells

doi: 10.1084/jem.20200839

Figure Lengend Snippet: NK cell killing of hiECs in the presence and absence of CD47. (A–E) WT, B2M − / − CIITA − / − , and B2M − / − CIITA − / − CD47 tg hiECs were challenged with primary NK cells (A) or NK cells from a cell line (B–E) using in vitro impedance assays. Graphs show mean ± SD and three independent replicates per group and time point; three different E:T ratios are shown. wt, wild-type.

Article Snippet: In some cases, cells were stimulated with mouse IL-2 (1 ng/ml; PeproTech) or stimulator cells were pretreated with anti-mouse Cd47 blocking antibody (clone MIAP301, rat IgG2a,κ; BioXCell) and effector cells with mouse FcR block (catalog no. 130–092-575, concentration 1:5; Miltenyi) for 120 min at 37°C.

Techniques: In Vitro

Journal: The Journal of Experimental Medicine

Article Title: The SIRPα–CD47 immune checkpoint in NK cells

doi: 10.1084/jem.20200839

Figure Lengend Snippet: Mechanistic interaction of CD47 with NK cell SIRPα. (A–E) B2M − / − CIITA − / − CD47 tg hiECs engaged with primary NK cells (A) or NK cells from a cell line (B–E) using in vitro impedance assays in the presence or absence of a specific blocking antibody against CD47 (clone B6.H12) or a blocking peptide against SIRPα. Graphs show mean ± SD and three independent replicates per group and time point; three different E:T ratios are shown.

Article Snippet: In some cases, cells were stimulated with mouse IL-2 (1 ng/ml; PeproTech) or stimulator cells were pretreated with anti-mouse Cd47 blocking antibody (clone MIAP301, rat IgG2a,κ; BioXCell) and effector cells with mouse FcR block (catalog no. 130–092-575, concentration 1:5; Miltenyi) for 120 min at 37°C.

Techniques: In Vitro, Blocking Assay

Journal: The Journal of Experimental Medicine

Article Title: The SIRPα–CD47 immune checkpoint in NK cells

doi: 10.1084/jem.20200839

Figure Lengend Snippet: CD47 overexpression using plasmid vector transfection. (A) CD47 overexpression in B2M − / − CIITA − / − hiPSCs was achieved by plasmid vector transfection, and out of this B2M − / − CIITA − / − CD47 tg (plasmid) hiPSC pool, 28 clones were picked and expanded. Clone 21 showed the highest CD47 expression. (B) Clone 21 iPSCs were differentiated into hiECs, and the CD47 expression level was assessed by flow cytometry (mean ± SD, four independent experiments). (C and D) hiEC clone 21 was challenged with IL-2–activated human primary NK cells (C) or human macrophages (D). Graphs show mean ± SD and three independent replicates per group and time point; three different E:T ratios are shown.

Article Snippet: In some cases, cells were stimulated with mouse IL-2 (1 ng/ml; PeproTech) or stimulator cells were pretreated with anti-mouse Cd47 blocking antibody (clone MIAP301, rat IgG2a,κ; BioXCell) and effector cells with mouse FcR block (catalog no. 130–092-575, concentration 1:5; Miltenyi) for 120 min at 37°C.

Techniques: Over Expression, Plasmid Preparation, Transfection, Clone Assay, Expressing, Flow Cytometry

Journal: The Journal of Experimental Medicine

Article Title: The SIRPα–CD47 immune checkpoint in NK cells

doi: 10.1084/jem.20200839

Figure Lengend Snippet: CD47 overexpression in K562 cells . (A) CD47 overexpression was achieved with lentiviral particles and the relative surface expression on K562 CD47 tg was assessed in flow cytometry (mean ± SD, three independent samples per group, Student’s t test). (B–D) CD3 − CD7 + CD56 + primary NK (B), NKL (C), or NK92 (D) cells were stimulated with IL-2 for 72 h before they were added to Fluc + K562 or K562 CD47 tg, and target cell killing was assessed in BLI assays (mean ± SD, three independent samples per group, Student’s t test). (E) Fluc + K562 cells were transplanted into NSG mice, which received adoptive transfer of IL-2–treated CD3 − CD7 + CD56 + primary NK cells. Cell survival was longitudinally monitored using BLI (five mice; each line represents one mouse). (F–G) Fluc + K562 CD47 tg cells were transplanted into NSG mice, and cell survival was longitudinally monitored using BLI (five mice per graph; each line represents one mouse). IL-2–treated CD3 − CD7 + CD56 + primary NK cells were used as effector cells without blocking CD47 (F) or with CD47 antibody block and FcR block (G; clone B6.H12). (H and I) Fluc + K562 CD47 tg cells were transplanted into NSG mice with (H) or without (I) CD47 antibody block but without transfer of human NK cells (three mice per graph; each line represents one mouse). (J) Fluc + K562 CD47 tg cells were transplanted into NSG mice that received IL-2–treated CD3 − CD7 + CD56 + primary NK cells (five mice; each line represents one mouse). Free Fab fragments of the CD47 antibody (clone B6.H12; H) were used to block CD47 interactions. (K) IL-2–treated NK92 effector cells were transferred in conjunction with the CD47 blocking antibody in mice transplanted with Fluc + K562 CD47 tg cells (five mice; each line represents one mouse).

Article Snippet: In some cases, cells were stimulated with mouse IL-2 (1 ng/ml; PeproTech) or stimulator cells were pretreated with anti-mouse Cd47 blocking antibody (clone MIAP301, rat IgG2a,κ; BioXCell) and effector cells with mouse FcR block (catalog no. 130–092-575, concentration 1:5; Miltenyi) for 120 min at 37°C.

Techniques: Over Expression, Expressing, Flow Cytometry, Adoptive Transfer Assay, Blocking Assay

Journal: The Journal of Experimental Medicine

Article Title: The SIRPα–CD47 immune checkpoint in NK cells

doi: 10.1084/jem.20200839

Figure Lengend Snippet: Inhibition of rhesus monkey NK cells and macrophages by B2M −/− CIITA −/− hiECs overexpressing rhesus CD47. (A) WT, B2M − / − CIITA − / − , and B2M − / − CIITA − / − CD47 tg hiECs were challenged with rhesus NK cells using in vitro impedance assays. Rhesus NK cells were used either unstimulated or stimulated with rhesus IL-2. Graphs show mean ± SD and three independent replicates per group and time point; three different E:T ratios are shown. (B) The rhesus CD47 tg was expressed in B2M − / − CIITA − / − rhCD47 hiECs, and expression by fluorescence is compared with primary rhesus ECs (representative histogram of two independent experiments). (C) B2M − / − CIITA − / − rhCD47 tg hiECs were challenged with unstimulated or rhesus IL-2–stimulated rhesus NK cells. Graphs show mean ± SD and three independent replicates per group and time point; three different E:T ratios are shown. (D) WT, B2M − / − CIITA − / − , and B2M − / − CIITA − / − rhCD47 tg hiECs were challenged with rhesus macrophages. Graphs show mean ± SD and three independent replicates per group and time point; three different E:T ratios are shown. wt, wild-type.

Article Snippet: In some cases, cells were stimulated with mouse IL-2 (1 ng/ml; PeproTech) or stimulator cells were pretreated with anti-mouse Cd47 blocking antibody (clone MIAP301, rat IgG2a,κ; BioXCell) and effector cells with mouse FcR block (catalog no. 130–092-575, concentration 1:5; Miltenyi) for 120 min at 37°C.

Techniques: Inhibition, In Vitro, Expressing, Fluorescence

Journal: Nature Communications

Article Title: Epithelial CD47 is critical for mucosal repair in the murine intestine in vivo

doi: 10.1038/s41467-019-12968-y

Figure Lengend Snippet: CD47 regulates mucosal wound healing in vivo. Utilizing a miniature video endoscope and biopsy scissors, 5–7 wounds were created in the dorsal aspect of the descending colon mucosa of anesthetized mice. a Digital measurement of wound surface area at 24 and 72 h post wounding revealed a striking impairment in wound closure in Cd47 −/− mice. Points represent the mean value within all wounds from individual mice. Data are representative of three independent experiments with five mice per group and are expressed as means ± SEM. *** p < 0.001; two-tailed Student’s t test. b In total, 10 µg of control antibody (IgG) or anti-CD47 antibody (miap301 or miap410) were injected into wound beds of wounds created 24 h previously in C57Bl/6 mice, resulting in substantial reduction of wound closure upon blockade of CD47. c Mice treated locally or systemically with anti-CD47 monoclonal antibodies miap301 or miap410 experienced less wound area reduction in comparison with IgG-treated controls. Points represent mean value within all wounds from an individual mouse. Data are representative of two independent experiments with five mice per group. Date are means ± SEM. *** p < 0.001; one-way ANOVA. Scale bars: 50 mm. Source data are provided as a Source Data file

Article Snippet: From BioXCell: Purified rat IgG2A (clone 2A3; BP0089), miap301 (BE0270), and miap410 (BE0283) at 10 μg/ml.

Techniques: In Vivo, Two Tailed Test, Control, Injection, Bioprocessing, Comparison

Journal: Nature Communications

Article Title: Epithelial CD47 is critical for mucosal repair in the murine intestine in vivo

doi: 10.1038/s41467-019-12968-y

Figure Lengend Snippet: CD47 is required for wound repair in cultures of primary epithelial monolayers. a Primary epithelial cell monolayers derived from CD47-expressing (CD47( + )) or CD47-deficient (CD47(−)) murine enteroids were scratch-wounded and monitored for closure. CD47(−) epithelial monolayers showed significant impairment in reduction of scratch-wound surface area at 24 h post scratch. Edges of scratch wounds are indicated by dashed lines. Scale bars = 50 μm. b Primary epithelial cell monolayers derived from human stem cell-derived colonoids were scratch-wounded and treated with 10 µg/ml of either IgG control antibody, function-blocking anti-CD47 antibody (clone B6H12), or non-blocking anti-CD47 antibody (clone 2D3), resulting in the inhibition of cell migration upon blockade of CD47. a – b Results are representative of three independent experiments with three replicates per treatment group. Data are means ± SEM. Significance determined by two-way ANOVA, ** p ≤ 0.01, *** p ≤ 0.001. Source data are provided as a Source Data file

Article Snippet: From BioXCell: Purified rat IgG2A (clone 2A3; BP0089), miap301 (BE0270), and miap410 (BE0283) at 10 μg/ml.

Techniques: Derivative Assay, Expressing, Control, Blocking Assay, Inhibition, Migration

Journal: Cancer immunology, immunotherapy : CII

Article Title: NECTIN4 regulates the cell surface expression of CD155 in non-small cell lung cancer cells and induces tumor resistance to PD-1 inhibitors.

doi: 10.1007/s00262-025-04079-z

Figure Lengend Snippet: Fig. 5 Overcoming anti- PD-1 antibody resistance in NECTIN4-overexpressing 4T1 cells by combination therapy with anti-TIGIT antibody. A, Schematic representation of the experiment: the subcutane- ous transplantation of 4T1 EV control or NECTIN4-overex- pressing cells, followed by treat- ment with anti-PD-1 antibody alone or in combination with anti-TIGIT antibody. The treat- ment was initiated on day 7 after tumor transplantation and administered every three days for a total of three times. Anti- PD-1 mAb, or IgG2a isotype were administered at a dose of 200 µg, and anti-TIGIT mAb or IgG1 isotype were administered at a dose of 150 µg. B, Graph showing tumor volume progres- sion after transplantation. C, Comparison of tumor weights in each group, measured post- excision. D, Photographs of tumors excised two days after the completion of treatment. E, Analysis of tumor-infiltrating lymphocytes (TILs) by flow cytometry. Experiments were conducted with five mice per group and the data are presented as the mean ± SD. All in vivo experiments were performed in duplicates, with similar results. Statistical analysis was con- ducted using one-way ANOVA followed by Tukey’s test (B, C) or Student’s t-test (E). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. EV, empty vec- tor; OE, overexpression

Article Snippet: One week after transplantation, tumorbearing mice were treated intraperitoneally with monoclonal antibodies (mAbs) as follows: 200 μg of anti-PD-1 mAb (RMP1-14, BioXcell, #BE0146), 150 μg of anti-TIGIT mAb (1G9, BioXcell, #BE0274), 200 μg of control IgG2a mAb (2A3, BioXcell, #BE0089), and 150 μg of control IgG1 mAb (MOPC-21, BioXcell, #BE0083).

Techniques: Transplantation Assay, Control, Comparison, Flow Cytometry, In Vivo, Over Expression

Journal: Clinical & Experimental Metastasis

Article Title: A new Neu—a syngeneic model of spontaneously metastatic HER2-positive breast cancer

doi: 10.1007/s10585-024-10289-z

Figure Lengend Snippet: NT2.5-LM responds to HER2-directed therapy. a 1 × 10 5 NT2.5-LM cells were injected into a mammary fat pad of NeuN mice. After surgical resection of NT2.5-LM tumor-bearing mice at 12 days post-injection (dpi), treatment with isotype or anti-HER2 monoclonal antibody (100 µg/mouse, 1x/week, intraperitoneal injection) began at 23 dpi (n = 12 per treatment group) and continued until survival endpoint at 70 dpi. b 1 × 10 5 NT2.5-LM cells were injected into a mammary fat pad of NeuN mice, tumors were surgically resected at 12 dpi, and anti-HER2 treatment (100 µg/mouse, 1x/week, intraperitoneal injection) began at 23 dpi (n = 10 per treatment group). Lungs were collected at 38 dpi. Three different levels were taken from formalin-fixed and paraffin-embedded lungs sectioned 100 µm apart. Slides were H&E stained, scanned, and analyzed using HALO to obtain summed lung metastasis counts and c percent tumor area over normal lung tissue. Two mice in the vehicle group were removed due to inconsistencies between HALO results and physical examination of H&E slides. Statistics used: Mantel-Cox Log-rank test for ( a ), Mann–Whitney U-test for ( b–c ), ns = not-significant, **p < 0.01

Article Snippet: Anti-HER2 monoclonal antibody (BioXCell, clone 7.16.4) and mouse IgG2a isotype vehicle antibody (BioXCell, clone C1.18.4) were dosed at 100 μg/mouse by intraperitoneal (i.p.) injection once a week for three weeks as described [ ].

Techniques: Injection, Staining, MANN-WHITNEY