Journal: Nature Cell Biology

Article Title: p16-dependent increase of PD-L1 stability regulates immunosurveillance of senescent cells

doi: 10.1038/s41556-024-01465-0

Figure Lengend Snippet: a , Imaging flow cytometry analysis of subcellular localization of p16 and PD-L1 staining within the AM population. Representative images of CD45 + PD-L1 + p16 + (row a), CD45 + PD-L1 − p16 + (row b) and CD45 + PD-L1 + p16 − (row c) cells. Bright field (BF); scale bars, 10 μm. Images are representative from three mice repeated independently with similar results. b , INs-seq of p16 + and p16 − AMs. GSEA of p16 + and p16 − AMs. Control ( n = 4) and inflamed (Infl; n = 4) samples were used. DESeq2 was used to derive gene fold changes for p16 + versus p16 − macrophages, controlling for treatment (LPS/PBS) as a covariant. NES, normalized enrichment score. c , d , Flow cytometry analysis of lung with the percentage of Foxp3 + Tregs within CD4 population ( c ) and the percentage of CD4 Foxp3 + Tregs expressing PD1 ( d ). Ctrl, control (PBS); Infl, inflamed (LPS). e , Experimental setup: the mice that were exposed daily to either PBS (Ctrl) or LPS (Infl) inhalations for 5 days received anti-PD1, anti-PD-L1 or matched IgG control as indicated, and the lungs and BAL were analysed 48 h after the last inhalation. FC, flow cytometry. f , g , Flow cytometry analysis of lung ( f ) or BAL ( g ) from the mice treated as in e . f , Percentage of p16 + PD-L1 + cells within CD45 + or AM cells. g , Percentage of CD8 + T cells positive for ICOS, CD25, CD44 and CD69. Two-sided Mann–Whitney U test ( c and d ) was used for statistical analysis. Error bars, mean ± s.e.m., *** P < 0.001. In f and g , one-way ANOVA was used for statistical analysis. Error bars, mean ± s.e.m., * P < 0.05, ** P < 0.01, *** P < 0.001. In b and d , control ( n = 5–9) and Infl ( n = 5–7) samples were used. In f and g , LPS + IgG ( n = 11), LPS + anti-PD1 ( n = 10) and LPS + anti-PD-L1 ( n = 10) samples were used. In c – g , experiments were repeated three times independently with similar results. Source numerical data are available in .

Article Snippet: For the immune checkpoint blockade treatment mice received intravenous injection of 200 μg anti-PD-L1 (Ichorbio, ICH1086), 200 μg anti-PD1 (Ichorbio, ICH1091) or 200 μg isotype control IgG2b (Ichorbio, ICH2243).

Techniques: Imaging, Flow Cytometry, Staining, Control, Expressing, MANN-WHITNEY

Journal: Nature Cell Biology

Article Title: p16-dependent increase of PD-L1 stability regulates immunosurveillance of senescent cells

doi: 10.1038/s41556-024-01465-0

Figure Lengend Snippet: (A) Experimental setup: mice were exposed daily to either PBS (Ctrl) or LPS inhalations for 5 days before analysis of mice lungs 48 h after the last inhalation. (B) Representative flow cytometry plots showing the gating strategy for p16 + PD-L1 + cells from immune cells (CD45 + ) and Alveolar Macrophage (AM) population. (C-E) Flow cytometry analysis of (C) CD45 + , (D) AM, and (E) p16 + PD-L1 + cells within CD45 + and AM populations in mice treated as described in A. (n = 6-10) (F) Experimental setup: mice exposed daily to either PBS (Ctrl) or LPS inhalations for 5 days received anti-PD-L1 or matched IgG control as indicated, and lungs and bronchoalveolar lavage (BAL) were analysed 48 h after the last inhalation. (G-H) Flow cytometry analysis of lung (G) or BAL (H) from mice treated as in scheme F. (G) Percentage of p16 + PD-L1 + cells within CD45 + or AM. (n = 5) ( H) Percentage of CD8 T cells positive for ICOS, CD25, CD69, and PD1 (n = 13-14) (I) mRNA level of IFN-γ cytokine from CD8 T cells in BAL. Values are relative to mice treated with IgG control (n = 3-4). (C-E, G-I) Two-sided Mann-Whitney test was used for statistical analysis. Error bars, mean ± SEM *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Experiments were repeated three times independently with similar results. Source numerical data are available in source data.

Article Snippet: For the immune checkpoint blockade treatment mice received intravenous injection of 200 μg anti-PD-L1 (Ichorbio, ICH1086), 200 μg anti-PD1 (Ichorbio, ICH1091) or 200 μg isotype control IgG2b (Ichorbio, ICH2243).

Techniques: Flow Cytometry, Control, MANN-WHITNEY

Journal: Nature Cell Biology

Article Title: p16-dependent increase of PD-L1 stability regulates immunosurveillance of senescent cells

doi: 10.1038/s41556-024-01465-0

Figure Lengend Snippet: a , Experimental setup included young or old mice that received an anti-PD-L1 or matched IgG control as indicated, and their lungs and blood were analysed 48 h after the last injection. FC, flow cytometry. b , c , Flow cytometry analysis of lungs from mice treated as in a , with a percentage of p16 + PD-L1 + cells within CD45 + or AM ( b ) and a percentage of CD8 + T cells positive for ICOS, CD25, CD44, CD69 and PD1 ( c ). d , Plasma levels of IFN-γ and IL-10. e , Experimental setup included mice that were exposed three times a week for 10 weeks to LPS (Infl) inhalations and anti-PD-L1 or matched IgG control as indicated, and their lungs were analysed 48 h after the last inhalation. Naive mice were the control group (Ctrl). f , Percentage of p16 + PD-L1 + cells within CD45 + or AM. g , Senescence-associated gene expression in the lungs of naive mice compared with the ones with chronic inflammation, treated with anti-PD-L1, or matched IgG control. For all experiments, statistical significance was calculated using one-way ANOVA; error bars, mean ± s.e.m. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. In a – d , young ( n = 7), old + IgG ( n = 7) and old + anti-PD-L1 ( n = 8) mice were used. In e – g , naive ( n = 6–7), Infl + IgG ( n = 7–10) and Infl + anti-PD-L1 ( n = 7–8) mice samples were used. Experiments were repeated three times independently with similar results. Source numerical data are available in .

Article Snippet: For the immune checkpoint blockade treatment mice received intravenous injection of 200 μg anti-PD-L1 (Ichorbio, ICH1086), 200 μg anti-PD1 (Ichorbio, ICH1091) or 200 μg isotype control IgG2b (Ichorbio, ICH2243).

Techniques: Control, Injection, Flow Cytometry, Expressing

Journal: Nature Cell Biology

Article Title: p16-dependent increase of PD-L1 stability regulates immunosurveillance of senescent cells

doi: 10.1038/s41556-024-01465-0

Figure Lengend Snippet: (A-D) Analysis of old mice treated with either anti-PD-L1 or isotype control (IgG). Young mice are the control group. ( A-B) Flow cytometry analysis of the lungs. (A) Representative flow cytometry plots showing the gating strategy for p16 + PD-L1 + cells from immune cells (CD45 + ) and Alveolar Macrophage (AM) population. (B) Percentage of NK cells positive for CD25, CD44, CD69, and PD1. (C) Plasma levels of IL6, IL17, and MCP-1. (D) Predicted age based on the epigenetic clock of the peripheral immune system in the blood. (E) Flow cytometry analysis of mice with chronic lung inflammation (Infl) treated with either anti-PD-L1 or IgG control. Naive mice were the control group (Ctrl). Percentage of CD8 T cells positive for ICOS, CD25, CD44, CD69, and PD1. (F) Scheme of anti-PD-L1 antibody treatment of senescent cells and its effect. Statistical significance was calculated using one-way ANOVA; n = 6 − 9, Error bars, mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, (B-D) young (n = 7), old + IgG (n = 7), old + anti-PD-L1 (n = 8); (E-H) naive (n = 7-8), Infl + IgG (n = 7-10), Infl + anti-PD-L1 (n = 7-8). Experiments were repeated three times independently with similar results. Source numerical data are available in source data.

Article Snippet: For the immune checkpoint blockade treatment mice received intravenous injection of 200 μg anti-PD-L1 (Ichorbio, ICH1086), 200 μg anti-PD1 (Ichorbio, ICH1091) or 200 μg isotype control IgG2b (Ichorbio, ICH2243).

Techniques: Control, Flow Cytometry

Journal: Molecular Therapy Oncology

Article Title: Promising immunotherapy targets: TIM3, LAG3, and TIGIT joined the party

doi: 10.1016/j.omton.2024.200773

Figure Lengend Snippet: Clinical trials on LAG3

Article Snippet: NCT03005782 , 2016 , 1 , REGN3767 , study of REGN3767 (anti-LAG3) with or without REGN2810 (anti-PD-1) in advanced cancers , Regeneron Pharmaceuticals (Tarrytown, NY, USA).

Techniques: Clinical Proteomics, Adjuvant, Expressing

Journal: Vaccines

Article Title: Macrophages as a Potential Immunotherapeutic Target in Solid Cancers

doi: 10.3390/vaccines11010055

Figure Lengend Snippet: Selected clinical trials of the TAM-targeting agents in combination with other therapeutic interventions.

Article Snippet: BI 754,091 (OSE Immunotherapeutics) , BI 754,091 (anti-PD1) , Solid tumors , Phase I , NCT03990233.

Techniques: Clinical Proteomics