Journal: Aging Cell

Article Title: Galectin‐9 high Neutrophils Exacerbate Radiation‐Induced Frailty

doi: 10.1111/acel.70448

Figure Lengend Snippet: GAL‐9 protein is an important regulatory molecule in neutrophil hyperactivity. (A) Intracellular GAL‐9 protein expression of circulating neutrophils in the sham group and 80 days post‐radiation. n = 6. (B) Circulating GAL‐9 protein level by ELISA kits. n = 8. (C, D) (C) Scheme and (D) the IFN‐γ expression of circulating neutrophils in the sham group under different conditions. n = 3. (E–M) (E) Scheme of the IFN‐γ and NETs expression of circulating neutrophils in the sham group by treatment with (F, H, K) circulating serum at 80 days post‐radiation and (F, I, L) culture supernatant of GAL‐9 high neutrophils. (F, J, M) IFN‐γ and NETs expression of GAL‐9 high neutrophils 80 days post‐radiation under different stimuli. n = 4–5. (N–Q) Assessment of the effect of GAL‐9 protein on the polarization of bone marrow macrophages under different conditions. (N) The scheme and the effect of (O) rmGAL‐9 protein, (P) culture supernatant of GAL‐9 high neutrophils, and (Q) circulating serum at 80 days post‐radiation were shown. n = 3. (R–S) Representative plots and statistics of bone marrow CD47 + neutrophils in the sham group and 80 days post‐radiation. n = 4. (T‐BB) Assessment of the reversal effect of GAL‐9 intervention in mice. (T) Scheme of the administration of anti‐GAL‐9 and rmGAL‐9 proteins in the local radiation group and the sham group, respectively. (U) Circulating GAL‐9 high neutrophils, bone marrow (V) CMP cells, (W) CLP cells, (X) non‐immune cells, (Y) macrophages, and (Z, AA) their polarization state and (BB) frailty index score were shown after different treatments in the local radiation group and the sham group. n = 3–5. Data are presented as mean ± SD; each dot represents an individual animal from at least 2–4 independent experiments that used male and female mice. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001. Statistical analyses were performed using unpaired Student's t ‐test (A, B, S, U), one‐way ANOVA (D, G–M, V–AA), and two‐way ANOVA (O–Q).

Article Snippet: Primary antibodies used were Gal‐9 (1:500, Abcam #ab69630), Myeloperoxidase (1:50, Abcam #ab90810), Histone H3 (1:1000, Abcam #ab281584), Ly6g + Ly6c (1:500, Abcam #ab25377), and CD47 (1:500, Abmart #T55251S).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Cell Reports Medicine

Article Title: Dual PD-1/IL-2Rα targeting restores CD8 + T cell fitness via STAT5/CD47 axis in SMARCA4-deficient NSCLC

doi: 10.1016/j.xcrm.2026.102633

Figure Lengend Snippet: PD-1/IL-2 bsAb protects CD8 + T cells from macrophage phagocytosis via STAT5-mediated upregulation of CD47 (A) Schematic of the Cleavage Under Targets and Tagmentation (CUT&Tag) assays workflow. (B) Distribution of STAT5 binding signals relative to transcription start sites (TSS) in CD8 + T cells treated with or without the PD-1/IL-2 bsAb. (C) Genomic annotation of differentially enriched STAT5 binding peaks in the bsAb-treated group. (D) KEGG pathway enrichment analysis of genes associated with STAT5 binding peaks. (E and F) Strategy and Venn diagram for identifying potential STAT5 downstream genes. (G) ChIP-qPCR analysis of STAT5 binding to the promoter regions of selected candidate genes ( n = 3). (H) Genome browser tracks showing STAT5 binding signals at the CD47 locus in control and PD-1/IL-2 bsAb-treated CD8 + T cells. (I) Schematic of the macrophage phagocytosis assay. (J) Representative confocal microscopy images showing macrophages (red) engulfing CD8 + T cells (green). Scale bars, 20 μm. (K) Flow cytometry quantification of the percentage of macrophages that had phagocytosed CD8 + T cells under the indicated conditions ( n = 3). Data are represented as mean ± SD (error bars) from biological replicates. Statistical analyses, n.s., no significance. Statistical significance was determined by unpaired Student’s t test or one-way ANOVA where appropriate.

Article Snippet: CD47 (E2V9V) Rabbit mAb , Cell Signaling Technology , Cat#36096.

Techniques: Binding Assay, ChIP-qPCR, Control, Phagocytosis Assay, Confocal Microscopy, Flow Cytometry

Journal: Cell Reports Medicine

Article Title: Dual PD-1/IL-2Rα targeting restores CD8 + T cell fitness via STAT5/CD47 axis in SMARCA4-deficient NSCLC

doi: 10.1016/j.xcrm.2026.102633

Figure Lengend Snippet: CD47 protects CD8 + T cells from macrophage clearance to boost antitumor immunity in SMARCA4-deficient NSCLC (A) Schematic of the adoptive T cell therapy experiment ( n = 8/group). (B) Representative in vivo bioluminescence images of mice from the indicated treatment groups at different time points. (C) Tumor growth curves, as measured by bioluminescence, for mice in each treatment group. (D) Individual tumor growth curves for mice in each treatment group. (E) Kaplan-Meier survival curves of mice from the four treatment groups. (F) Quantification by flow cytometry of donor-derived CD45.2 + CD8 + T cells among total tumor-infiltrating lymphocytes. (G) Representative flow cytometry plots for donor-derived CD45.2 + CD8 + T cells expressing the exhaustion markers PD-1, TIGIT, and TIM-3. (H) Quantification of the percentage of CD45.2 + CD8 + T cells expressing PD-1, TIGIT, and TIM-3. (I) The production of TNF-α by donor-derived CD45.2 + CD8 + T cells. (J) The production of IFN-γ by donor-derived CD45.2 + CD8 + T cells. (K) Representative immunofluorescence images of tumor sections. White: CD8, green: CD47, red: F4/80. Scale bars, 70 μm. (L) Schematic model depicting the proposed mechanism of action. Data are represented as mean ± SD (error bars) from biological replicates. Statistical analyses, n.s., no significance. Statistical significance was determined by one-way ANOVA.

Article Snippet: CD47 (E2V9V) Rabbit mAb , Cell Signaling Technology , Cat#36096.

Techniques: In Vivo, Flow Cytometry, Derivative Assay, Expressing, Immunofluorescence

Journal: Cell Reports Medicine

Article Title: Dual PD-1/IL-2Rα targeting restores CD8 + T cell fitness via STAT5/CD47 axis in SMARCA4-deficient NSCLC

doi: 10.1016/j.xcrm.2026.102633

Figure Lengend Snippet: PD-1/IL-2 bsAb protects CD8 + T cells from macrophage phagocytosis via STAT5-mediated upregulation of CD47 (A) Schematic of the Cleavage Under Targets and Tagmentation (CUT&Tag) assays workflow. (B) Distribution of STAT5 binding signals relative to transcription start sites (TSS) in CD8 + T cells treated with or without the PD-1/IL-2 bsAb. (C) Genomic annotation of differentially enriched STAT5 binding peaks in the bsAb-treated group. (D) KEGG pathway enrichment analysis of genes associated with STAT5 binding peaks. (E and F) Strategy and Venn diagram for identifying potential STAT5 downstream genes. (G) ChIP-qPCR analysis of STAT5 binding to the promoter regions of selected candidate genes ( n = 3). (H) Genome browser tracks showing STAT5 binding signals at the CD47 locus in control and PD-1/IL-2 bsAb-treated CD8 + T cells. (I) Schematic of the macrophage phagocytosis assay. (J) Representative confocal microscopy images showing macrophages (red) engulfing CD8 + T cells (green). Scale bars, 20 μm. (K) Flow cytometry quantification of the percentage of macrophages that had phagocytosed CD8 + T cells under the indicated conditions ( n = 3). Data are represented as mean ± SD (error bars) from biological replicates. Statistical analyses, n.s., no significance. Statistical significance was determined by unpaired Student’s t test or one-way ANOVA where appropriate.

Article Snippet: CD47 Polyclonal Ab , Proteintech , Cat#20305-1-AP; RRID: AB_10732838.

Techniques: Binding Assay, ChIP-qPCR, Control, Phagocytosis Assay, Confocal Microscopy, Flow Cytometry

Journal: Cell Reports Medicine

Article Title: Dual PD-1/IL-2Rα targeting restores CD8 + T cell fitness via STAT5/CD47 axis in SMARCA4-deficient NSCLC

doi: 10.1016/j.xcrm.2026.102633

Figure Lengend Snippet: CD47 protects CD8 + T cells from macrophage clearance to boost antitumor immunity in SMARCA4-deficient NSCLC (A) Schematic of the adoptive T cell therapy experiment ( n = 8/group). (B) Representative in vivo bioluminescence images of mice from the indicated treatment groups at different time points. (C) Tumor growth curves, as measured by bioluminescence, for mice in each treatment group. (D) Individual tumor growth curves for mice in each treatment group. (E) Kaplan-Meier survival curves of mice from the four treatment groups. (F) Quantification by flow cytometry of donor-derived CD45.2 + CD8 + T cells among total tumor-infiltrating lymphocytes. (G) Representative flow cytometry plots for donor-derived CD45.2 + CD8 + T cells expressing the exhaustion markers PD-1, TIGIT, and TIM-3. (H) Quantification of the percentage of CD45.2 + CD8 + T cells expressing PD-1, TIGIT, and TIM-3. (I) The production of TNF-α by donor-derived CD45.2 + CD8 + T cells. (J) The production of IFN-γ by donor-derived CD45.2 + CD8 + T cells. (K) Representative immunofluorescence images of tumor sections. White: CD8, green: CD47, red: F4/80. Scale bars, 70 μm. (L) Schematic model depicting the proposed mechanism of action. Data are represented as mean ± SD (error bars) from biological replicates. Statistical analyses, n.s., no significance. Statistical significance was determined by one-way ANOVA.

Article Snippet: CD47 Polyclonal Ab , Proteintech , Cat#20305-1-AP; RRID: AB_10732838.

Techniques: In Vivo, Flow Cytometry, Derivative Assay, Expressing, Immunofluorescence

Journal: Clinical and Experimental Medicine

Article Title: The role of macrophages and cytokines in the occurrence and development of MDS

doi: 10.1007/s10238-026-02106-6

Figure Lengend Snippet: Mechanism of action of anti-CD47 antibody.The CD47 molecule on the cell surface evades macrophage phagocytosis by binding to SIRPα. Overexpression of CD47 by cancer cells masks prephagocytic signals to evade macrophage phagocytosis.When CD47 is blocked, the prephagocytic signal on the surface of tumor cells is exposed and binds to the lowdensitylipoprotein receptor-related protein (LRP) on the surface of macrophages, thereby achieving phagocytosis and removal of tumor cells

Article Snippet: .On the other hand, anti-CD47 antibodies can block the binding of CD47 to macrophages while exposing a pro-phagocytosis signal, which is a prephagocytic cell signal [ ].

Techniques: Binding Assay, Over Expression