Fig. 1
Engineering of 20.1 mAb reveals lack of BTN3A1 clustering in Vγ9Vδ2 activation. (a) Schematic representing the putative effects of increasing 20.1 inter-Fab distance on BTN3A1 membrane organization. (b) Schematic representing key features of modified mAb constructs with hinge-regions lengthened using Glycine-Serine linkers. (c) Activation of G115-TCR expressing Jurkat Jrt-3.5 cells after co-incubation with DaudiΔAAVS1 cells pre-incubated with 20.1 hinge variants at increasing concentrations or control (Buffer). CD69 expression as a marker of T cell activation was assessed by flow-cytometry (gated on isotype antibody). Means + SD and non-linear regression with variable slope—four parameters (n = 3). (d) EC50 of agonism and 95% confidence intervals (CI) for 20.1 hinge variants was calculated by non-linear regression with variable slope—four parameters (R2 = 0.9706, 0.9920, 0.9744, 0.8868, 0.9843, 0.9857, respectively). # = significant difference in EC50 value determined by non-overlapping CI.
