human primary cd8 cytotoxic t cells  (ATCC)

Journal: Journal of Nanobiotechnology

Article Title: CD8a antibody-functionalized biomimetic red blood cell membrane ectosomes delivering C646 reverse CD8⁺ T Cell exhaustion via H3K18la histone delactylation in gastric cardia adenocarcinoma

doi: 10.1186/s12951-025-03957-z

Figure Lengend Snippet: Bioinformatics Analysis Identifies Key Factors in Histone Lactylation Modification. Note: ( A ) Schematic workflow of bioinformatics analysis for identifying key factors; ( B ) Volcano plot of DEGs in tumor tissues and adjacent normal tissues from dataset GSE2685 (Normal = 8, Tumor = 22); ( C ) Heatmap showing the correlation of co-expression module genes with tumor and normal tissues, with each cell displaying the correlation coefficient and p -value; ( D ) Venn diagram illustrating the intersection of Blue module genes, DEGs, CD8 + T cell-related genes, and "Histone lactylation"-related genes; ( E ) Box plot of p300 differential expression; ( F ) The tSNE distribution map of EP300 in various cell types in the scRNA-seq data

Article Snippet: Human CD8+ T cells were isolated from peripheral blood mononuclear cells (hPBMCs, PCS-800–011, ATCC, USA) using the CD8 MicroBead Kit (130–045–201, Miltenyi Biotec, USA).

Techniques: Modification, Expressing, Quantitative Proteomics

Journal: Journal of Nanobiotechnology

Article Title: CD8a antibody-functionalized biomimetic red blood cell membrane ectosomes delivering C646 reverse CD8⁺ T Cell exhaustion via H3K18la histone delactylation in gastric cardia adenocarcinoma

doi: 10.1186/s12951-025-03957-z

Figure Lengend Snippet: p300 Regulates Histone Lactylation in CD8 + T Cells. Note: ( A ) Schematic representation of the experimental design, showing the workflow for detecting CD8 + T cells treated with lactate, p300 inhibitors, or activators. ( B–C ) WB analysis of PKla levels in CD8 + T cells over time (B) and under varying lactate concentrations ( C ). ( D–E ) WB analysis of the time-dependent ( D ) and dose-dependent ( E ) changes in H3K18la and H3K9la expression in CD8 + T cells following lactate treatment. *** p < 0.001, ** p < 0.01, and * p < 0.05 compared to the 0-h or untreated lactate group. ( F ) WB analysis of H3K18la and H3K9la expression in CD8 + T cells following p300 knockdown, activation, or inhibition. ( G ) ELISA detection of IFN-γ levels in the supernatant of CD8 + T cells across different treatment groups. (H) FCM analysis of GZMB expression in CD8 + T cells. ( I ) FCM analysis of CD8 + T cell proliferation. ( J ) LDH release assay showing the cytotoxic effects of CD8 + T cells on MKN-45 and SNU1 cells. In panels ( F–J ), * p < 0.05, ** p < 0.01, and *** p < 0.001 compared to the control group; # p < 0.05, ## p < 0.01, and ### p < 0.001 compared to the Lactate group; & p < 0.01 compared to the Lactate + sh-NC group. All cell-based experiments were performed in triplicate

Article Snippet: Human CD8+ T cells were isolated from peripheral blood mononuclear cells (hPBMCs, PCS-800–011, ATCC, USA) using the CD8 MicroBead Kit (130–045–201, Miltenyi Biotec, USA).

Techniques: Expressing, Knockdown, Activation Assay, Inhibition, Enzyme-linked Immunosorbent Assay, Lactate Dehydrogenase Assay, Control

Journal: Journal of Nanobiotechnology

Article Title: CD8a antibody-functionalized biomimetic red blood cell membrane ectosomes delivering C646 reverse CD8⁺ T Cell exhaustion via H3K18la histone delactylation in gastric cardia adenocarcinoma

doi: 10.1186/s12951-025-03957-z

Figure Lengend Snippet: In Vivo and In Vitro Biocompatibility and Targeting Validation of NVEs. Note: ( A ) Schematic illustration of in vitro targeting and biocompatibility experiments of NVEs; ( B ) Hemolysis analysis of NVEs, NVEs@C646, and CD8a-NVEs@C646 after incubation with red blood cells (PC: positive control); ( C ) CD8 + T cell viability assessed using the CCK-8 assay after 48-h incubation with varying concentrations of NVEs; ( D ) Confocal microscopy and quantitative analysis of NVEs@C646 and CD8a-NVEs@C646 uptake in CD8 + T cells (Scale bars = 25 μm); (E) FCM analysis of the uptake efficiency of NVEs@C646 and CD8a-NVEs@C646 in CD8 + T cells; (F) Schematic illustration of the subcutaneous tumor model in mice and in vivo injection of NVEs; ( G ) FCM analysis of the percentage of CM-Dil-labeled CD8 + T cells in blood, spleen, tumor tissue, and TdLNs at different time points post-injection (* p < 0.05 compared with the NVEs@C646 group); ( H ) FCM analysis of CD3 + CD8 − T cells binding with CM-Dil-labeled NVEs recovered from blood, spleen, tumor tissue, and TdLNs at different time points; ( I ) Quantification of CD8 + T cell numbers 48 h after NVEs injection. Cell-based experiments were performed in triplicate. Animal experiments included nine mice per group, with three mice per time point

Article Snippet: Human CD8+ T cells were isolated from peripheral blood mononuclear cells (hPBMCs, PCS-800–011, ATCC, USA) using the CD8 MicroBead Kit (130–045–201, Miltenyi Biotec, USA).

Techniques: In Vivo, In Vitro, Biomarker Discovery, Incubation, Positive Control, CCK-8 Assay, Confocal Microscopy, Injection, Labeling, Binding Assay

Journal: Journal of Nanobiotechnology

Article Title: CD8a antibody-functionalized biomimetic red blood cell membrane ectosomes delivering C646 reverse CD8⁺ T Cell exhaustion via H3K18la histone delactylation in gastric cardia adenocarcinoma

doi: 10.1186/s12951-025-03957-z

Figure Lengend Snippet: CD8a-NVEs@C646 Facilitates Histone Delactylation Modification in CD8 + T Cells. Note: ( A ) Schematic workflow of RNA-seq and ChIP-seq experiments for CD8 + T cells treated with CD8a-NVEs@C646; ( B–C ) ChIP-seq analysis showing signals at TSS regions in PBS-treated (n = 3) and CD8a-NVEs@C646-treated groups (n = 3); ( D ) RNA-seq volcano plot illustrating significantly upregulated and downregulated genes in the CD8a-NVEs@C646-treated group compared to PBS (n = 3); ( E ) Venn diagram of overlapping genes from ChIP-seq and RNA-seq analyses related to CD8 + T cells; ( F ) ChIP analysis of p300 and H3K18la enrichment at the PDCD1 promoter region; ( G ) The protein interaction between p300 and H3K18la was detected by co-IP assay; ( H ) RT-qPCR analysis of PDCD1 mRNA expression in CD8 + T cells following CD8a-NVEs@C646 treatment; ( I ) WB analysis of PDCD1 and H3K18la protein expression levels in CD8 + T cells treated with CD8a-NVEs@C646. * p < 0.05, *** p < 0.001 compared to the PBS or sh-NC group; experiments were conducted in triplicate

Article Snippet: Human CD8+ T cells were isolated from peripheral blood mononuclear cells (hPBMCs, PCS-800–011, ATCC, USA) using the CD8 MicroBead Kit (130–045–201, Miltenyi Biotec, USA).

Techniques: Modification, RNA Sequencing, ChIP-sequencing, Co-Immunoprecipitation Assay, Quantitative RT-PCR, Expressing

Journal: Journal of Nanobiotechnology

Article Title: CD8a antibody-functionalized biomimetic red blood cell membrane ectosomes delivering C646 reverse CD8⁺ T Cell exhaustion via H3K18la histone delactylation in gastric cardia adenocarcinoma

doi: 10.1186/s12951-025-03957-z

Figure Lengend Snippet: CD8a-NVEs@C646-Mediated Regulation of PDCD1 Enhances the Tumor-Killing Ability of CD8 + T Cells. Note: ( A ) Schematic diagram of the experimental design: CD8 + T cells were transduced with PDCD1 overexpression lentivirus and treated with CD8a-NVEs@C646, followed by co-culture with MKN-45/SNU1 cells; ( B ) RT-qPCR analysis of PDCD1 mRNA expression in CD8 + T cells across different groups; ( C ) WB analysis of PDCD1 and H3K18la protein expression in CD8 + T cells across different groups; ( D ) ELISA detection of IFN-γ levels in the supernatant of CD8 + T cells across different groups; ( E ) FCM analysis of GZMB expression in CD8 + T cells across different groups; ( F ) FCM analysis of CD8 + T cell proliferation activity; ( G ) LDH release assay to evaluate the cytotoxicity of CD8 + T cells on MKN-45 cells; ( H ) FCM analysis of apoptosis in MKN-45 cells across different groups. * indicates p < 0.05 compared to the PBS + vector group, *** indicates p < 0.001, # indicates p < 0.05 compared to the CD8a-NVEs@C646 + vector group, ### indicates p < 0.001. All cellular experiments were repeated three times

Article Snippet: Human CD8+ T cells were isolated from peripheral blood mononuclear cells (hPBMCs, PCS-800–011, ATCC, USA) using the CD8 MicroBead Kit (130–045–201, Miltenyi Biotec, USA).

Techniques: Transduction, Over Expression, Co-Culture Assay, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Activity Assay, Lactate Dehydrogenase Assay, Plasmid Preparation

Journal: Journal of Nanobiotechnology

Article Title: CD8a antibody-functionalized biomimetic red blood cell membrane ectosomes delivering C646 reverse CD8⁺ T Cell exhaustion via H3K18la histone delactylation in gastric cardia adenocarcinoma

doi: 10.1186/s12951-025-03957-z

Figure Lengend Snippet: CD8a-NVEs@C646 Enhances CD8 + T Cell-Mediated Anti-Tumor Immunity. Note: ( A ) Schematic diagram of the experimental design, illustrating the establishment of the GCA orthotopic transplantation tumor model and treatment regimen; ( B ) In vivo fluorescence imaging of tumors in mice from different treatment groups and quantitative analysis of signal intensity over time; ( C ) Photographic images of tumor tissues from different treatment groups; ( D ) Tumor weight statistics for each group; ( E–F ) Immunohistochemistry and TUNEL staining showing cell proliferation and apoptosis in tumor tissues from each group (Scale bars = 50 μm); ( G ) Quantification of the proportion of infiltrating CD8 + T cells in tumor tissues from each group; ( H-J ) Expression levels of IFN-γ, TNFα, and GZMB in CD8 + T cells within tumor tissues from each group; ( K ) Proportion of PD-1 + CD8 + T cells in tumor tissues from each group. Six mice per group; * indicates p < 0.05 compared to the NS group, and # indicates p < 0.05 compared to the αPD-1 group

Article Snippet: Human CD8+ T cells were isolated from peripheral blood mononuclear cells (hPBMCs, PCS-800–011, ATCC, USA) using the CD8 MicroBead Kit (130–045–201, Miltenyi Biotec, USA).

Techniques: Transplantation Assay, In Vivo, Fluorescence, Imaging, Immunohistochemistry, TUNEL Assay, Staining, Expressing