Journal: iScience

Article Title: A transfer learning framework to elucidate the clinical relevance of altered proximal tubule cell states in kidney disease

doi: 10.1016/j.isci.2024.109271

Figure Lengend Snippet: Characterization of altered proximal tubule cells (A) UMAP representation of the PT compartment from AKI and control patients. (B) Relative abundance of PT1 and PT2 clusters in AKI and control patients. (C) Expression of known injury marker genes in PT1 and PT2 clusters. (D and E) Gene-weighted density of HAVCR1 and VCAM1. (F) Enrichment score calculated by gene set enrichment analysis using Reactome pathway database (positive enrichment means an enrichment in PT2 cluster). (G) Pathway activity in PT1 and PT2 clusters (inferred from PROGENy). (H) Collagen, extracellular matrix (ecm) proteoglycan (pg) and glycoprotein (gp) scores in PT1 and PT2 clusters. (I) Potential of heat-diffusion for affinity-based trajectory embedding (PHATE) dimension reduction projecting pathway enrichment estimated by GSVA for each cell type. (J and K) Representative immunostainings of HAVCR1 (J) and VCAM-1 (K) proteins in fibrotic area. ∗∗∗p < 0.001 ∗∗∗∗p < 0.0001.

Article Snippet: mouse monoclonal anti-human HAVCR1 , clone 219211 , MAB1750; RRID: AB_2116559 ; RD Systems.

Techniques: Control, Expressing, Marker, Activity Assay, Diffusion-based Assay

Journal: iScience

Article Title: A transfer learning framework to elucidate the clinical relevance of altered proximal tubule cell states in kidney disease

doi: 10.1016/j.isci.2024.109271

Figure Lengend Snippet:

Article Snippet: mouse monoclonal anti-human HAVCR1 , clone 219211 , MAB1750; RRID: AB_2116559 ; RD Systems.

Techniques: Control, Software

Journal: International immunopharmacology

Article Title: Dominant negative TGFβ receptor II and truncated TIM3 enhance the antitumor efficacy of CAR-T-cell therapy in prostate cancer.

doi: 10.1016/j.intimp.2023.110807

Figure Lengend Snippet: Fig. 1. Expression of TIM3 and TGFβRII and their ligands in prostate cancer: (A) Expression of GAL9 in prostate cancer and normal prostate tissues in the GEPIA 2 database; (B) Expression of TGF-β in prostate cancer and normal prostate tissues in the GEPIA 2 database; (C) Correlation between the expression of TIM3 and TGFβRII, GAL9, and TGF-β in the GEPIA 2 database; (D) IHC of GAL9 in prostate cancer in the HPA database; (E) IHC of TGF-β in prostate cancer in the HPA database; (F) The left panel shows unsupervised graph-based clustering of all samples visualized by UMAP delineated by cell type, and the right panel shows the expression of TIM3 and TGFβRII in different cell populations in the single-cell sequencing results; (G) Expression of TIM3 and TGFβRII in T cells in the single-cell sequencing results (the left panel is a scatter plot, and the right panel is a violin plot); (H) Coexpression of TIM3 and TGFβRII in T cells in the single-cell sequencing results, where 1 (blue) represents T cells that coexpress TIM3 and TGFβRII, and 0 (gray) represents other T cells.

Article Snippet: Untransduced (UTD) T cells, PSMA-CAR-T cells, and DT-PSMA-CAR- L. Tang et al. International Immunopharmacology 124 (2023) 110807 L. Tang et al. International Immunopharmacology 124 (2023) 110807 L. Tang et al. International Immunopharmacology 124 (2023) 110807 T cells were co-cultured with prostate cancer cells at ratios of 1:1, 5:1, and 10:1 for 24 h.The T-cell culture medium was supplemented with appropriate concentrations (0 ng/ml, 5 ng/ml, 20 ng/ml, and 50 ng/ml) of TGF-β (P01137, Suzhou Novoprotein Scientific) and/or (0 μg/ml, 5 μg/ml, 10 μg/ml, and 20 μg/ml) TIM3 activating monoclonal antibodies (MAB23651, R&D Systems) from the day before coculture to the end of coculture.

Techniques: Expressing, Sequencing

Journal: International immunopharmacology

Article Title: Dominant negative TGFβ receptor II and truncated TIM3 enhance the antitumor efficacy of CAR-T-cell therapy in prostate cancer.

doi: 10.1016/j.intimp.2023.110807

Figure Lengend Snippet: Fig. 2. Preparation and phenotypic characterization of DT-PSMA-CAR-T cells: (A) Gene structure of PSMA-CAR and DT-PSMA-CAR; (B) Expansion of the UTD group, PSMA-CAR-T group, and DT-PSMA-CAR-T group T cells 14 days after electroporation; (C) The positivity rate of CAR in the PSMA-CAR-T group and DT-PSMA-CAR-T group, after magnetic bead sorting, assessed by flow cytometry; (D) Expression of TIM3 and TGFβRII on the three groups of T cells assessed by flow cytometry before performing in vitro killing assays; (E) Expression of CD4 and CD8 on the three groups of T cells assessed by flow cytometry before performing in vitro killing assays; (F) Expression of CD45RO on the three groups of T cells assessed by flow cytometry before performing in vitro killing assays.

Article Snippet: Untransduced (UTD) T cells, PSMA-CAR-T cells, and DT-PSMA-CAR- L. Tang et al. International Immunopharmacology 124 (2023) 110807 L. Tang et al. International Immunopharmacology 124 (2023) 110807 L. Tang et al. International Immunopharmacology 124 (2023) 110807 T cells were co-cultured with prostate cancer cells at ratios of 1:1, 5:1, and 10:1 for 24 h.The T-cell culture medium was supplemented with appropriate concentrations (0 ng/ml, 5 ng/ml, 20 ng/ml, and 50 ng/ml) of TGF-β (P01137, Suzhou Novoprotein Scientific) and/or (0 μg/ml, 5 μg/ml, 10 μg/ml, and 20 μg/ml) TIM3 activating monoclonal antibodies (MAB23651, R&D Systems) from the day before coculture to the end of coculture.

Techniques: Electroporation, Flow Cytometry, Expressing, In Vitro

Journal: International immunopharmacology

Article Title: Dominant negative TGFβ receptor II and truncated TIM3 enhance the antitumor efficacy of CAR-T-cell therapy in prostate cancer.

doi: 10.1016/j.intimp.2023.110807

Figure Lengend Snippet: Fig. 4. Analysis of the resistance of DT-PSMA-CAR-T cells to immune suppression caused by TIM3 activation in vitro was performed by tumor lysis assays: (A and B) UTD group, PSMA-CAR-T group, and DT-PSMA-CAR-T group T cells were cocultured with LNCAP cells in the presence of 10 μg/ml TIM3 activating monoclonal antibody; (C and D) Under an effector-to-target ratio of 5:1, the three groups of T cells were cocultured with LNCAP cells in the presence of different concentrations of TIM3 activating monoclonal antibody; (E) Cytokine secretion was detected in the supernatant of all experimental groups in (C).

Article Snippet: Untransduced (UTD) T cells, PSMA-CAR-T cells, and DT-PSMA-CAR- L. Tang et al. International Immunopharmacology 124 (2023) 110807 L. Tang et al. International Immunopharmacology 124 (2023) 110807 L. Tang et al. International Immunopharmacology 124 (2023) 110807 T cells were co-cultured with prostate cancer cells at ratios of 1:1, 5:1, and 10:1 for 24 h.The T-cell culture medium was supplemented with appropriate concentrations (0 ng/ml, 5 ng/ml, 20 ng/ml, and 50 ng/ml) of TGF-β (P01137, Suzhou Novoprotein Scientific) and/or (0 μg/ml, 5 μg/ml, 10 μg/ml, and 20 μg/ml) TIM3 activating monoclonal antibodies (MAB23651, R&D Systems) from the day before coculture to the end of coculture.

Techniques: Activation Assay, In Vitro, Lysis

Journal: International immunopharmacology

Article Title: Dominant negative TGFβ receptor II and truncated TIM3 enhance the antitumor efficacy of CAR-T-cell therapy in prostate cancer.

doi: 10.1016/j.intimp.2023.110807

Figure Lengend Snippet: Fig. 5. Analysis of the resistance of DT-PSMA-CAR-T cells to immune suppression caused by TIM3 and TGFβRII activation in vitro was performed by tumor lysis assays: (A and B) UTD group, PSMA-CAR-T group, and DT-PSMA-CAR-T group T cells were cocultured with LNCAP cells in the presence of 20 ng/ml TGF-β and 10 μg/ml TIM3 activating monoclonal antibody; (C and D) Under an effector-to-target ratio of 5:1, the three groups of T cells were cocultured with LNCAP cells in the presence of different combinations of TGF-β and TIM3 activating monoclonal antibody; (E) Cytokine secretion was detected in the supernatant of all experimental groups in (C).

Article Snippet: Untransduced (UTD) T cells, PSMA-CAR-T cells, and DT-PSMA-CAR- L. Tang et al. International Immunopharmacology 124 (2023) 110807 L. Tang et al. International Immunopharmacology 124 (2023) 110807 L. Tang et al. International Immunopharmacology 124 (2023) 110807 T cells were co-cultured with prostate cancer cells at ratios of 1:1, 5:1, and 10:1 for 24 h.The T-cell culture medium was supplemented with appropriate concentrations (0 ng/ml, 5 ng/ml, 20 ng/ml, and 50 ng/ml) of TGF-β (P01137, Suzhou Novoprotein Scientific) and/or (0 μg/ml, 5 μg/ml, 10 μg/ml, and 20 μg/ml) TIM3 activating monoclonal antibodies (MAB23651, R&D Systems) from the day before coculture to the end of coculture.

Techniques: Activation Assay, In Vitro, Lysis

Journal: International immunopharmacology

Article Title: Dominant negative TGFβ receptor II and truncated TIM3 enhance the antitumor efficacy of CAR-T-cell therapy in prostate cancer.

doi: 10.1016/j.intimp.2023.110807

Figure Lengend Snippet: Fig. 6. DT-PSMA-CAR-T cells can inhibit the growth of NSG mouse xe nografts in an environment activated by TIM3 and TGFβRII: (A) The timeline of animal experiments: 1 × 106 GAL9- PSMA-PC3 cells were subcutaneously injected; two weeks later, 5 × 106 UTD T cells, PSMA-CAR-T cells, or DT- PSMA-CAR-T cells were infused per mouse via the tail vein; and biolumi nescence imaging was performed weekly; (B) The location and tumor burden of the xenografts displayed by bioluminescence imaging; (C) Quanti tative results of tumor burden obtained by Aniview100.

Article Snippet: Untransduced (UTD) T cells, PSMA-CAR-T cells, and DT-PSMA-CAR- L. Tang et al. International Immunopharmacology 124 (2023) 110807 L. Tang et al. International Immunopharmacology 124 (2023) 110807 L. Tang et al. International Immunopharmacology 124 (2023) 110807 T cells were co-cultured with prostate cancer cells at ratios of 1:1, 5:1, and 10:1 for 24 h.The T-cell culture medium was supplemented with appropriate concentrations (0 ng/ml, 5 ng/ml, 20 ng/ml, and 50 ng/ml) of TGF-β (P01137, Suzhou Novoprotein Scientific) and/or (0 μg/ml, 5 μg/ml, 10 μg/ml, and 20 μg/ml) TIM3 activating monoclonal antibodies (MAB23651, R&D Systems) from the day before coculture to the end of coculture.

Techniques: Injection, Imaging