Journal: bioRxiv

Article Title: Targeting Langerhans cells using a modular mannosylated nucleic acid-based vaccine platform

doi: 10.1101/2025.06.04.657560

Figure Lengend Snippet: Delivery of gp100 on mannosylated HJ scaffolds into moLCs for antigen presentation. A ) Reaction scheme for gp100-functionalization of Q2 and Q4 with NHS-DBCO ( 1 ), followed by SPAAC reaction to azide-modified ( pink ) Val-Cit-PAB-linked ( yellow) gp100 ( red ) ( 2 ). B ) Schematic representation of gp100-conjugated HJs employed for antigen presentation. C ) Visual representation of the antigen-presentation assay using moLCs incubated with gp100-conjugated HJs to facilitate receptor-mediated endocytosis ( I ), followed by release of gp100 in the lysosomes ( II ) and presentation of gp100 on HLA-A2 ( III ). Subsequently, moLCs were co-cultured with wildtype- or gp100 Jurkat cells to verify antigen presentation by T cell activation. D ) IL-2 concentration (pg/mL) in the co-culture media by ELISA. Results shown from four independent human donors were expressed by the mean ± SD. Statistical analysis was conducted by a one-way ANOVA followed by a Tukey’s post hoc test. E ) Antigen-presentation assay comparing HJ-3xTriMan-1xgp100 assembled with either a cleavable linker ( Val-Cit ) or a non-cleavable linker. Results shown from three independent human donors were expressed by the mean ± SD. Statistical analysis was conducted by a two-way t test. p-value * = <0.05, ** = <0.01, *** = <0.001, ****= <0.0001.

Article Snippet: Antigenic peptide gp100 154-162 (sequence: KTWGQYWQV, specificity: HLA-A*02:01, GENAXXON bioscience) was conjugated to the oligonucleotide strands in a 2-step one-pot reaction.

Techniques: Immunopeptidomics, Modification, Incubation, Cell Culture, Activation Assay, Concentration Assay, Co-Culture Assay, Enzyme-linked Immunosorbent Assay