Journal: NPJ Vaccines
Article Title: Antigen cross-presentation potentiating cancer vaccine adjuvant for T cell expansion and synergy with anti-PD-1
doi: 10.1038/s41541-026-01376-1
Figure Lengend Snippet: A The vector map of recombinant protein constructs: FMS-like tyrosine kinase-3 ligand extracellular domain (Flt3L/F), Vibrio vulnificus flagellin B (FlaB/B). The fusion protein was generated by combining F and B with B position at the C-terminal (FB) or the N-terminal (BF). B Characterization of the recombinant proteins by sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE) and subsequent Western blot analysis using mouse anti-FlaB serum or rabbit anti-Flt3L antibody. C Determination of TLR5-dependent NF-κB stimulating activity of B, FB, and BF in various protein concentrations using HEK-Blue™ hTLR5 cells ( n = 3 biological replicates). D Total cell number and the number of cell cluster formations of BMDCs after 9 days of cultures supplemented with B, F, and FB. Floating cells were collected and counted using a Vi-Cell Blue machine ( n = 7 biological replicates), while cell clusters were visualized with an EMOS5000 machine ( n = 5 biological replicates). E Representative flow histogram of CD103 expression on CD11c + MHCII hi population and quantification data of CD103 + DCs generation by recombination protein in a dose-dependent manner using BMDCs ( n = 3 biological replicates). F Representative levels of CFSE in CD8 + T cells from cultures with the indicated DCs, the individual peaks of CFSE dilution gated, and quantitative proliferation index of CD8 + T cell. For all sections: **** P < 0.0001; *** P < 0.001; ** P < 0.01; * P < 0.05.
Article Snippet: To assess the TLR5-stimulating activities of the recombinant fusion proteins, we evaluated the TLR5-dependent NF-κB-stimulating activity of the recombinant protein using HEK-BlueTM hTLR5 cells (hκb-help-5, InvivoGen, USA) and HEK-BlueTM Detection (hb-det2, InvivoGen, USA) assay systems following the manufacturer’s instructions.
Techniques: Plasmid Preparation, Recombinant, Construct, Generated, SDS Page, Western Blot, Activity Assay, Expressing
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