Journal: Journal of Nanobiotechnology
Article Title: Engineering BiTE-inspired IPSC-exosomes to potentiate CAR-T cell therapy against lung cancer
doi: 10.1186/s12951-026-04242-3
Figure Lengend Snippet: Functional characterization and T-cell bridging activity. ( a , b ) Binding affinity analysis of engineered exosomes (MIEXO, PIEXO, BIEXO) to MSLN (a) and PD-1 ( b ) via indirect ELISA. Exosomes were added to PD-1 or MSLN-coated plates and detected via an HRP-conjugated anti-His-tag antibody. ( c , d ) Flow cytometric analysis of engineered exosomes (100 µg/mL) binding to PD-1 + T cells ( c ) and LLC-MSLN cells ( d ), with His-tag mean fluorescence intensity (MFI) quantification (right panel). ( e , f ) Confocal microscopy analysis of BIEXO targeting to PD-1 + T cells ( e ) and LLC-MSLN cells ( f ). ( g ) Representative confocal microscopy images and corresponding quantification showing BIEXO-mediated bridging between PD-1 + T cells and LLC-MSLN cells. ( h ) FRET analysis of dose-dependent bridging efficiency. ( i , j ) Prevention of PD-L1-induced T-cell exhaustion measured by LDH release ( n = 3). ( k , l ) Experimental design (k) for ex vivo culture of CD8 + T cells isolated from mouse spleen, and flow cytometric analysis of TCF-1 + Tpex cell expansion in PD-1 + CD8 + T cells following pre-treatment with the AhR antagonist CH-223,191 and BIEXO@IPA stimulation ( l ). ( m ) In vitro growth inhibition of LLC-MSLN cells following treatment. ( n ) Enhanced cytotoxicity of TILs isolated from LLC-MSLN tumor-bearing mice against LLC-MSLN cells following BIEXO@IPA treatment. Data are presented as mean ± SD. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test (c, d, j, m, n). * p < 0.05 , ** p < 0.01 , *** p < 0.001 , **** p < 0.0001
Article Snippet: Recombinant human PD-1 protein (5 μg/mL, R&D Systems, 1086-PD) was used for capture.
Techniques: Functional Assay, Activity Assay, Binding Assay, Indirect ELISA, Fluorescence, Confocal Microscopy, Ex Vivo, Isolation, In Vitro, Inhibition