Journal: Science Advances

Article Title: Inflammatory arthritis irAE may represent a unique autoimmune disease primarily driven by T cells but likely not autoantibodies

doi: 10.1126/sciadv.aea4262

Figure Lengend Snippet: ( A to G ) PBMCs from the patients with irAE were cultured in the plate coated with anti-CD3 and anti-CD28 (10 μg/ml) in the presence of IgG1 isotype control or anti–human IL-6R (50 μg/ml), anti–human IL-12p40 (50 μg/ml), and anti–human IFNAR1 (50 μg/ml) for 3 days; n = 9. (A) Expression of CD38 and CD127 on CD8 + T cells. Right: Percentage of CD38 + CD127 − CD8 + T cells. (B and C) CD8 + T cells activated for 5 days were restimulated with PMA, ionomycin, and monensin for 5 hours. (B) Expression of granzyme B and IFN-γ. Right: Percentage of granzyme B + IFN-γ + CD8 + T cells. (C) Expression of perforin and IFN-γ. Right: Percentage of perforin + IFN-γ + CD8 + T cells. [(D) and (E)] MFIs of GluCy5 (D), TMRM, and MTDR (E) in CD8 + T cells were presented. [(F) and (G)] CD4 + T cells activated for 5 days were restimulated with PMA, ionomycin, and monensin for 5 hours. (F) Expression of IL-21 and IL-2. Right: Percentage of IL-21 + IL-2 + CD4 + T cells. (G) Expression of CD38 and CXCR5 on CD4 + T cells. Right: Percentage of CD38 + CXCR5 − CD4 + T cells. Data in graphs represent mean ± SEM. Significance was tested paired Student’s t test [(A) to (G)].

Article Snippet: PBMCs from patients with irAE were thawed and rested in a complete medium for 1 hour and then activated with plate-coated anti–human CD3 and anti–human CD28 (10 μg/ml) with IgG1 isotype control (Bio X Cell, catalog no. CP174) or a combination of anti–human IL-6R 50 μg/ml; Bio X Cell, catalog no. SIM0014), anti–human IL-12p40 (50 μg/ml; Bio X Cell, catalog no. SIM0020), and anti–human IFNAR1 (50 μg/ml; Bio X Cell, catalog no. SIM0022) for 3 days.

Techniques: Cell Culture, Control, Expressing

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: SMARCA4 deficiency impairs CD8 + T cell function and confers resistance to PD-1 blockade in NSCLC (A) Progression-free survival (PFS) of eight patients with SMARCA4-deficient (BRG1-deficient) NSCLC treated with ICIs. (B) Representative axial CT images of a patient (P6) with SMARCA4-deficient NSCLC before and after ICIs treatment. (C) Change in the sum of target lesion diameters for six evaluable patients with SMARCA4-deficient NSCLC from baseline to first radiographic assessment. (D) Schematic of the experimental design for the orthotopic lung cancer model ( n = 8/group). (E) Representative in vivo bioluminescence images of mice from the indicated treatment groups at different time points. (F) Quantification of total bioluminescence flux from mice in each group over time. (G) Kaplan-Meier survival curves of mice from the four treatment groups. (H) Individual tumor growth curves, as measured by bioluminescence, for mice in each treatment group. (I) UMAP visualization of major immune cell populations within the TME, analyzed by flow cytometry. (J) Quantitative comparison of the frequencies of indicated immune cell lineages between SMARCA4-WT and -KD tumors. (K and L) Representative flow cytometry plots showing the production of IFN-γ and TNF-α by tumor-infiltrating CD8 + T cells. (M) Quantification of the frequencies of IFN-γ + and TNF-α + cells among tumor-infiltrating CD8 + T cells. 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- or two-way ANOVA where appropriate.

Article Snippet: CD8 + T cell dependency was assessed by co-administering a depleting anti-human CD8α antibody (BioXcell, 15 μg/mouse) with PD-1/IL-2 bsAb in the SMARCA4-KD model. Depletion (initiated one day before PD-1/IL-2 bsAb and maintained twice weekly) was confirmed by flow cytometry (>90% reduction in circulating human CD3 + CD8 + T cells).

Techniques: Cell Function Assay, In Vivo, Flow Cytometry, Comparison

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: Attenuated IL-2/STAT5 signaling and an enhanced exhaustion phenotype in CD8 + T cells within SMARCA4-deficient tumors (A) Schematic of the workflow for transcriptomic profiling of CD8 + T cells. (B) Volcano plot displaying differentially expressed genes in CD8 + T cells from SMARCA4-KD versus WT tumors. (C–E) Pathway enrichment analyses of genes downregulated in CD8 + T cells from SMARCA4-KD tumors, including Gene Ontology (GO) terms, KEGG pathways, and Reactome pathways. (F) The gene set enrichment analysis (GSEA) plot. (G) Correlation matrix (pie chart) showing the association between IL-2 receptor subunits expression and key T cell exhaustion marker genes in tumor-infiltrating CD8 + T cells. (H) Radar plot comparing the normalized expression levels of genes encoding IL-2 receptor subunits and exhaustion markers in CD8 + T cells. (I) Quantification by flow cytometry of the expression frequencies of PD-1, TIGIT, and TIM-3 on tumor-infiltrating CD8 + T cells. (J and K) Representative multiplex immunofluorescence (mIF) images of SMARCA4-WT and -KD tumor sections stained for PanCK (cyan), CD8 (green), GZMB (white), PD-1 (red), TIGIT (orange), and TIM-3 (yellow). Scale bars, 20 μm. 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: CD8 + T cell dependency was assessed by co-administering a depleting anti-human CD8α antibody (BioXcell, 15 μg/mouse) with PD-1/IL-2 bsAb in the SMARCA4-KD model. Depletion (initiated one day before PD-1/IL-2 bsAb and maintained twice weekly) was confirmed by flow cytometry (>90% reduction in circulating human CD3 + CD8 + T cells).

Techniques: Expressing, Marker, Flow Cytometry, Multiplex Assay, Immunofluorescence, Staining

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: SMARCA4 loss in tumor cells attenuates CD8 + T cell function via NF-κB-mediated suppression of ICAM1 (A) Flow cytometry analysis of surface expression of the exhaustion markers PD-1, TIGIT, and TIM-3 on human CD8 + T cells following co-culture. (B and C) Representative flow cytometry plots showing the production of IFN-γ and TNF-α and the surface expression of IL-2Rα (CD25) by CD8 + T cells following co-culture. (D) Quantification of the frequencies of IFN-γ + , TNF-α + , and IL-2Rα + cells among co-cultured CD8 + T cells. (E) Integrated single-nucleus RNA sequencing (snRNA-seq) analysis comparing IL2-STAT5 signaling activity. y axis: IL2-STAT5 signaling score. (F and G) Incoming and outgoing signaling patterns between major cell types in the TME, as inferred from snRNA-seq. (H) Specific cell-cell communication network illustrating the ICAM signaling pathway from tumor cells to CD8 + T cells in patients with SMARCA4-WT NSCLC. (I) Correlation analysis between SMARCA4 and ICAM1 mRNA expression in TCGA cohorts. (J) Immunohistochemistry staining and quantification of ICAM1 protein expression in tumor tissues from SMARCA4-WT ( n = 10) and -deficient ( n = 10) NSCLC patients. (K) Schematic illustrating the proposed link between SMARCA4 deficiency and impaired NF-κB activation. (L) Immunoblot analysis of ICAM1 and p65 protein levels in SMARCA4-WT H2122 cells treated with the NF-κB inhibitor PTDC or vehicle control. (M) ChIP-qPCR analysis showing NF-κB (p65) binding to a specific site within the ICAM1 promoter in SMARCA4-WT H2122 cells ( n = 3). (N) Dual-luciferase reporter assay in SMARCA4-WT H2122 cells ( n = 3). Data are represented as mean ± SD (error bars) from biological replicates. Statistical analyses, n.s., no significance, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001. Statistical significance was determined by unpaired Student’s t test or one-way ANOVA where appropriate.

Article Snippet: CD8 + T cell dependency was assessed by co-administering a depleting anti-human CD8α antibody (BioXcell, 15 μg/mouse) with PD-1/IL-2 bsAb in the SMARCA4-KD model. Depletion (initiated one day before PD-1/IL-2 bsAb and maintained twice weekly) was confirmed by flow cytometry (>90% reduction in circulating human CD3 + CD8 + T cells).

Techniques: Cell Function Assay, Flow Cytometry, Expressing, Co-Culture Assay, Cell Culture, RNA Sequencing, Activity Assay, Immunohistochemistry, Staining, Activation Assay, Western Blot, Control, ChIP-qPCR, Binding Assay, Luciferase, Reporter 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: The PD-1/IL-2 bsAb exerts potent anti-tumor efficacy in SMARCA4-deficient models (A) Schematic of the therapeutic experiment in humanized mice ( n = 6/group). (B) Representative images of excised tumors from each treatment group at the endpoint. (C) Tumor growth curves of individual mice in the indicated treatment groups. (D) Kaplan-Meier survival curves of mice from the four treatment groups. (E) Quantification by flow cytometry of tumor-infiltrating CD3 + CD8 + T cells. (F) Quantification of the percentage of tumor-infiltrating CD8 + T cells expressing PD-1, TIGIT, and TIM-3. (G) Frequency of polyfunctional (TNF-α + IFN-γ + ) CD8 + T cells among tumor-infiltrating lymphocytes. (H) Schematic of the humanized patient-derived xenograft (PDX) model ( n = 5/group). (I) Representative images of excised PDX tumors from each treatment group. (J) Tumor growth curves of individual PDX-bearing mice. (K) Kaplan-Meier survival curves of PDX-bearing mice from the three treatment groups. (L) Quantification of tumor-infiltrating CD3 + CD8 + T cells in PDX tumors. (M) Quantification of the percentage of CD8 + T cells from PDX tumors expressing PD-1, TIGIT, and TIM-3. (N) Frequency of polyfunctional (TNF-α + IFN-γ + ) CD8 + T cells in PDX tumors. (O) Representative multiplex immunofluorescence (mIF) images of PDX tumor sections from different treatment groups, stained for PanCK (cyan), CD8 (green), GZMB (white), PD-1 (red), TIGIT (orange), and TIM-3 (yellow). Scale bars, 40 μm. 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 or two-way ANOVA where appropriate.

Article Snippet: CD8 + T cell dependency was assessed by co-administering a depleting anti-human CD8α antibody (BioXcell, 15 μg/mouse) with PD-1/IL-2 bsAb in the SMARCA4-KD model. Depletion (initiated one day before PD-1/IL-2 bsAb and maintained twice weekly) was confirmed by flow cytometry (>90% reduction in circulating human CD3 + CD8 + T cells).

Techniques: Flow Cytometry, Expressing, Derivative Assay, Multiplex Assay, Immunofluorescence, Staining

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: STAT5 activation mediates the therapeutic effect of the PD-1/IL-2 bsAb in SMARCA4-deficient NSCLC (A) Representative flow cytometry plots showing the expression of the exhaustion markers PD-1, TIGIT, and TIM-3 on activated human CD8 + T cells. (B) Quantification of the mean fluorescence intensity (MFI) of the exhaustion markers PD-1, TIGIT, and TIM-3 on CD8 + T cells from the experiment in (A) ( n = 3). (C) Representative flow cytometry plots showing the production of IFN-γ and TNF-α by CD8 + T cells under the conditions described in (A). (D) Quantification of the frequencies of IFN-γ + and TNF-α + cells among CD8 + T cells ( n = 3). (E) Schematic of the adoptive T cell transfer experiment ( n = 8/group). (F) Representative in vivo bioluminescence images of mice from the indicated groups at different time points. (G) Tumor growth curves, as measured by bioluminescence, for mice in each treatment group. (H) Kaplan-Meier survival curves of mice from the four treatment groups. (I) Quantification of the frequency of donor-derived CD45.2 + CD8 + T cells among total tumor-infiltrating lymphocytes. (J) Flow analysis of donor-derived CD45.2 + CD8 + T cells expressing PD-1, TIGIT, and TIM-3. (K–M) Frequency of IFN-γ + (K–L) and TNF-α + (M) cells among donor-derived CD45.2 + CD8 + T cells. 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 or two-way ANOVA where appropriate.

Article Snippet: CD8 + T cell dependency was assessed by co-administering a depleting anti-human CD8α antibody (BioXcell, 15 μg/mouse) with PD-1/IL-2 bsAb in the SMARCA4-KD model. Depletion (initiated one day before PD-1/IL-2 bsAb and maintained twice weekly) was confirmed by flow cytometry (>90% reduction in circulating human CD3 + CD8 + T cells).

Techniques: Activation Assay, Flow Cytometry, Expressing, Fluorescence, In Vivo, Derivative 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: CD8 + T cell dependency was assessed by co-administering a depleting anti-human CD8α antibody (BioXcell, 15 μg/mouse) with PD-1/IL-2 bsAb in the SMARCA4-KD model. Depletion (initiated one day before PD-1/IL-2 bsAb and maintained twice weekly) was confirmed by flow cytometry (>90% reduction in circulating human CD3 + CD8 + T cells).

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: CD8 + T cell dependency was assessed by co-administering a depleting anti-human CD8α antibody (BioXcell, 15 μg/mouse) with PD-1/IL-2 bsAb in the SMARCA4-KD model. Depletion (initiated one day before PD-1/IL-2 bsAb and maintained twice weekly) was confirmed by flow cytometry (>90% reduction in circulating human CD3 + CD8 + T cells).

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

Journal: bioRxiv

Article Title: Differential CD8+ T/NK cell-mediated reduction of HIV-1 replication after combination of ART with TIGIT or KLRG1 blockade in humanized mice

doi: 10.1101/2025.09.11.675307

Figure Lengend Snippet: (A): Representative schematic representation of the different antibodies used in the study. Variable chain regions are highlighted in green for those recognizing KLRG1 and in orange for those recognizing TIGIT. The human IgG1 heavy chain region was consistent across all antibodies. (B): Dose response analysis of intracellular expression of CD107a and IFNγ in NK from a selected PWH after culture with increasing concentrations of aTIGIT (orange), aKLRG1 (green) and bispecific (purple) mAbs. (C): Fold change in proportions to isotype treated-group of CD107a+ IFNγ-cells within CD56+ CD16+ NK cells from n=7 PWH, in response to the selected 5μg/ml of the different antibody treatment groups. (D): Representative flow cytometry plots showing IFNγ versus CD107a expression in CD8+ T cells after MLR assay under different individual or bispecific aTIGIT and aKLRG1 mAbs. (E): Fold change in proportions of CD107a+ IFNγ-(left) and CD107a+ IFNγ+ (right) in CD8+ T cells from n=6 HIV negative donors, in the presence of the different mentioned mAbs. In panel C and E, data are presented as Box and Whiskers plots showing median values, with minimum and maximum error bars. Statistical significance was calculated using a using a Wilcoxon t test. *p<0.05.

Article Snippet: A humanized IgG1 isotypic mAb (BioXCell) mAbs was used as control for in vitro and in vivo experiments.

Techniques: Expressing, Flow Cytometry, Mlr Assay

Journal: bioRxiv

Article Title: Differential CD8+ T/NK cell-mediated reduction of HIV-1 replication after combination of ART with TIGIT or KLRG1 blockade in humanized mice

doi: 10.1101/2025.09.11.675307

Figure Lengend Snippet: (A): Spearman correlations between proportions of degranulation marker CD107a in CD8+ T cells in peripheral blood (PB; left) or spleen (SP; right) and HIV-1 pVL post-ATI in aKLRG1 mAb treated hBLT mice. (B): Spearman correlation between proportions of CD107a+ IFNγ+ TNFα-CD8+ T cells in SP and HIV-1 pVL post-ATI. Dots from mice treated with aKLRG1 and isotype mAbs are colored in green and blue, respectively (C): Spearman correlation between proportions of CD107a+ IFNγ+ TNFα-CD8+ T cells and proportions of CD107a-IFNγ+ CD8+ T cells located in SP of hBLT mice treated with aKLRG1 mAbs. Dots from mice treated with aKLRG1 and isotype mAbs are colored in green and blue, respectively (D): Representative confocal microscopy image showing expression of IFNγ (green), p24 (white) and CD8 (red) in the spleen from representative hBLT mice treated with an Isotypic IgG (left) and aKLRG1 (right) mAbs. (E): Number of CD8+ T cells expressing IFNγ normalized to total number of CD8+ T cells in SP from the different hBLT treatment groups (F, G): Spearman correlation between number of CD8+ IFNγ+ T cells into the white pulp areas versus number of infected p24+ cells (G) or the proportions of NKG2C-CD57+ in PB (G) at the end of experiment in the hBLT mice group treated with aKLRG1 mAb. Dots from mice treated with aKLRG1 and isotype mAbs are colored in green and blue, respectively. In panel E data are presented as Box and Whiskers plots showing median values, with minimum and maximum error bars. Statistical significance was calculated using a Spearman correlation test or a Mann-Whitney test. *p<0.05; **p<0.01.

Article Snippet: A humanized IgG1 isotypic mAb (BioXCell) mAbs was used as control for in vitro and in vivo experiments.

Techniques: Marker, Confocal Microscopy, Expressing, Infection, MANN-WHITNEY