Fig. 1: CXCR4 is upregulated on dendritic cells by therapeutic proteins in proportion to their immunogenic potential.

a Representative flow cytometry dot plots for CXCR4⁺ moDC (CD11c⁺HLA-DR⁺) after 24 h stimulation of moDC with media (“untreated”), LPS 1 μg/mL (“LPS”), or KLH 5 μg/mL + LPS 100 pg/mL (“KLH”). “CXCR4 FMO” represents a control sample stained with all antibody-fluorophore pairs except CXCR4 APC. Supplementary Fig. 1 shows the gating strategy for moDC (CD11c⁺HLA-DR⁺). b Frequency (%) of CXCR4⁺ moDCs (CD11c⁺HLA-DR⁺) for a representative donor (#236) after 24 h stimulation with media or KLH (5, 20, 50, 100, or 500 μg/mL) plus 100 pg/mL LPS. c Frequency (%) of CXCR4⁺ moDCs (CD11c⁺HLA-DR⁺) for a representative donor (#014) after 24 h stimulation with media (untreated), LPS (1 μg/mL), or 5 μg/mL tocilizumab, ATR-107, HuA33, or KLH (with 100 pg/mL LPS as in a). In b, c, treatments were tested in triplicate, each dot represents a technical replicate and error bars are mean ± SEM. Statistical significance between treatments was determined by a two-tailed unpaired student’s t-test (α = 0.05). Exact p-values are reported unless p > 0.05, i.e., not significant. Estimated effect sizes (Cohen’s d) and results for one-way ANOVA with Dunnett’s multiple comparisons test are in Supplementary Data.