Journal: Materials Today Bio

Article Title: Activated T-cell membrane-derived nanocargoes displaying multi-immune checkpoints for enhanced cancer immunotherapy

doi: 10.1016/j.mtbio.2025.102702

Figure Lengend Snippet: Association between intratumoural STING signature and ICs expression. (A–H) Representative IHC image and quantification of the expression level of STING and ICs including PD-L1/PD-1, GAL-9/TIM-3 and GAL-3/LAG-3 in lung cancer patients. (I) Experimental timeline and indicated treatment groups (three intravenous injections, diABZI: 0.75 mg per kg (body weight), n = 6 mice). (J) Statistical analysis tumor volume on day 13 of mice with CT-26 tumors receiving the indicated treatments. (K–O) Representative flow cytometric densities of total immune cells (CD45 + ), T lymphocytes (CD3 + ), DCs (CD11c + MHC-II + ), Macrophages (CD11b + F4/80 + ) and NK cells (CD3 − CD49b + ) in the tumors receiving diABZI or PBS at three days after the last injections (n = 6 mice). (P–R) Representative flow cytometric quantification of the expression level of PD-1, TIM-3 and LAG-3 in tumor-infiltrating CD3 + T cells of CT-26-bearing mice receiving diABZI or PBS at three days after the last injections (n = 6 mice). All data are presented as the mean ± S.D. The p values were determined by two-tailed Student's t -test for (B–H) and (J–R).

Article Snippet: 66220-1-Ig), LAG-3 Polyclonal antibody (Proteintech:16616-1-AP), and Recombinant Anti-TIM-3 antibody (Abcam: EPR22241 ), PD-L1 Rabbit Polyclonal Antibody (Abiowell: cat.no.

Techniques: Expressing, Two Tailed Test

Journal: Materials Today Bio

Article Title: Activated T-cell membrane-derived nanocargoes displaying multi-immune checkpoints for enhanced cancer immunotherapy

doi: 10.1016/j.mtbio.2025.102702

Figure Lengend Snippet: Screening activated T-cell with high expression of ICs. (A) Diagram of activated T cells with multiple ICs screening. (B–D) Representative flow cytometric histograms (left) and curves (right) of the expression level of ICs including PD-1 (B), LAG-3 (C) and TIM-3 (D) of activated T cells from day 0–8 of the T cell screening assay, gated on CD3 + T cells, CD4 + T cells, CD8 + T cells (n = 3 independent experiments), MFI, mean fluorescence intensity. (E) The KEGG pathway enrichment analysis of all types of membrane-associated genes that are upregulated in activated T cells compared with un-activated T cells. (F) Heatmap showing the scaled expression of membrane-associated genes in activated T cells compared with un-activated T cells. All data are presented as the mean ± s.d.

Article Snippet: 66220-1-Ig), LAG-3 Polyclonal antibody (Proteintech:16616-1-AP), and Recombinant Anti-TIM-3 antibody (Abcam: EPR22241 ), PD-L1 Rabbit Polyclonal Antibody (Abiowell: cat.no.

Techniques: Expressing, Screening Assay, Fluorescence, Membrane

Journal: Materials Today Bio

Article Title: Activated T-cell membrane-derived nanocargoes displaying multi-immune checkpoints for enhanced cancer immunotherapy

doi: 10.1016/j.mtbio.2025.102702

Figure Lengend Snippet: The preparation and characterizations of AM-dLNPs. (A)Transmission electron microscopy images of non-activated T cells membrane vesicles (M) and activated T cells membrane vesicles (AM). Scale bars, 100 nm. (B) Western blot analysis of CD3 protein in activated T cell (AT) and AM. (C, D) The average hydrodynamic diameter and surface zeta potential of AM and M as measured by dynamic light scattering (n = 3 independent experiments). (E) Rescue of ICs-mediated T-cell exhaustion by AM in cocultures of IFN-γ-pretreated (20 ng/ml, 24 h) 4T1-OVA/Luci cells and OT-1 T cells at an effector: target ratio of 5:1. 4T1-OVA/Luci cells death in the cocultures was evaluated by luciferase reporter assay system (n = 3 independent experiments). (F) Representative flow cytometric percentage of IFN-γ+, Perforin + and Granzyme B + (CzmB) in OT-1 T cells after co-incubation with IFN-γ-pretreated 4T1-OVA/Luci cells and diABZI, dLNPs, M-dLNPs, AM-LNPs, AM-dLNPs or PBS for 24h at an effector: target ratio of 5:1. (G–J) The average hydrodynamic diameter (G, H), polydispersity (I) and surface zeta potential (J) of T-cell membrane (TM), dLNPs, M-dLNPs, AM-LNPs and AM-dLNPs, as measured by dynamic light scattering (n = 3 independent experiments). (K) Western blot analysis of CD3, PD-1, LAG-3, and TIM-3 in dLNPs, M-dLNPs, AM-LNPs and AM-dLNPs. (L) Expression of PD-L1 on AM-derived nanocargoes was determined by Flow NanoAnalyzer. (M) Expression of TIM-3 on AM-derived nanocargoes was determined by Flow NanoAnalyzer. (N) Transmission electron microscopy images of dLNPs, M-dLNPs, AM-LNPs and AM-dLNPs. Scale bars, 200 nm. (O) High-performance liquid chromatography analysis of diABZI concentrations in dLNPs, M-dLNPs, AM-LNPs and AM-dLNPs (n = 3 independent experiments). dLNPs: diABZI-loaded liposomes, M-dLNPs: non-actived T-cell membrane-decorated diABZI-loaded liposomes, AM-LNPs: actived T-cell membrane-decorated empty liposomes, AM-dLNPs: actived T-cell membrane-decorated diABZI-loaded liposomes. All the data are presented as the mean ± S.D. The p values were determined by one-way ANOVA with Tukey's post-test for (E).

Article Snippet: 66220-1-Ig), LAG-3 Polyclonal antibody (Proteintech:16616-1-AP), and Recombinant Anti-TIM-3 antibody (Abcam: EPR22241 ), PD-L1 Rabbit Polyclonal Antibody (Abiowell: cat.no.

Techniques: Transmission Assay, Electron Microscopy, Membrane, Western Blot, Zeta Potential Analyzer, Luciferase, Reporter Assay, Incubation, Expressing, Derivative Assay, High Performance Liquid Chromatography, Liposomes

Journal: Materials Today Bio

Article Title: Activated T-cell membrane-derived nanocargoes displaying multi-immune checkpoints for enhanced cancer immunotherapy

doi: 10.1016/j.mtbio.2025.102702

Figure Lengend Snippet: AM-dLNPs induced potent anti-tumor immune responses. (A–B) Representative flow cytometry plots (left) and statistical analysis (right) of T cell exhaustion markers PD-1 (A) and LAG-3 (B) on CD3 + T cells across different treatment groups by flow cytometry (n = 5 mice). (C–D) Representative flow cytometric plots and frequencies of Perforin (C) and Gzm B (D) that generated by tumor-infiltrating CD8 + T cells harvested on day 3 from bearing CT-26 mice after the last injection of the three-injection regimen. (n = 5 mice). (E) Representative IF images of tumor sections obtained from mice receiving three injections of the indicated treatments. Gzm B (green) and Perforin (red) were visualized using fluorescence-labelled antibodies. The nuclei were stained with DAPI (blue). Scale bars, 25 μm. (F) Representative flow cytometry plots and statistical analysis of naïve T cells (CD44 − CD62L + ), central memory T cells (CD44 + CD62L + , TCM), effector memory T cells (CD44 + CD62L − , TEM) and terminal exhausted T cells (TE X ) within the TME across different treatment groups (n = 5 mice). (G, H) Representative flow cytometric frequencies of CD86 + DCs (G) and NK cells (H) in spleen harvested on day 3 from CT-26-bearing mice after the last injection of the three-injection regimen (n = 5 mice). All the data are expressed as mean ± S.D. The p values were determined by one-way ANOVA with Tukey's post-test for (A–D), (F) and (G, H). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: 66220-1-Ig), LAG-3 Polyclonal antibody (Proteintech:16616-1-AP), and Recombinant Anti-TIM-3 antibody (Abcam: EPR22241 ), PD-L1 Rabbit Polyclonal Antibody (Abiowell: cat.no.

Techniques: Flow Cytometry, Generated, Injection, Fluorescence, Staining

Journal: Materials Today Bio

Article Title: Developing immune-regulatory materials using immobilized monosaccharides with immune-instructive properties

doi: 10.1016/j.mtbio.2020.100080

Figure Lengend Snippet: Flow cytometry data are shown as a heat map summarizing the expression profile of three surface markers (CD86, CD40, and CD274) on DCs after incubation on immobilized monosaccharide libraries in the presence of a TLR4 ligand (LPS). Unstimulated (immature) DCs were used as a negative control. All changes are expressed as a percentage of expression levels in mature (LPS-stimulated) DCs. A reduction in CD40 expression was observed in most conditions with the highest reduction in fucose-contained libraries compared to LPS alone. CD86 expression was higher in a number of conditions (particularly mixers of Gal1 and Gal2); however, these did not reach statistical significance. CD274 (PD-L1) expression showed a significant increase in Man1 and Gal2 combinations. Data are shown as mean ± SD of three independent donors where ∗ p <0.05, ∗∗ p <0.01, ∗∗∗ p <0.001, and ∗∗∗∗ p <0.0001.

Article Snippet: DCs were then incubated with labeled antibodies CD274 (APC clone REA1197), CD40 PE (clone HB14), and CD86 FITC (clone FM95) and isotype-matched mouse antibody controls (all from Miltenyi Biotec) for 20 min in the dark at 4 °C.

Techniques: Flow Cytometry, Expressing, Incubation, Negative Control