Fig. 3

Nb-TriTE enhances T cell activation and function compared with Nb-BiTE. Nb-TriTE-induced T cells upregulation of CD25 (A), CD69 (B) and CD107a (C) in comparison with controls as analyzed by flow cytometry in the presence of HepG2-FAP cells at a 2 μg/mL protein concentration after 24 h and an E:T ratio of 10:1. D Proliferation of T cells in response to Nb-TriTE in comparison with other groups was measured by flow cytometry in the presence of HepG2-FAP cells after 5 days and an E:T ratio of 10:1, shown as a percentage divided. E Representative histograms of CD25, CD69, and CD107a expression and proliferation indices are shown for each group. F–G Subpopulation analysis based on CD62L and CD45RA expression induced by the Nb-TriTE or Nb-BiTE after 14 days. CD45RA⁻CD62L⁻cells represent effector memory T (TEM) cells, CD45RA⁻CD62L⁺cells represent central memory T (TCM) cells, and CD45RA⁺CD62L⁺cells represent naive T cells. Pie chart for CD62L/D45RA expression (left). (H-I) Levels of IL-2 and IFN-γ secretion in the presence of HepG2-FAP cells upon addition of Nb-TriTE, Nb-BiTE or Irrelevant ctr and Nb-TriTE + hPD-1 at equimolar concentrations. (J-K) An ELISPOT assay was performed to measure the IFN-γ response of specific T cells for each group. All data represent the mean ± standard deviation from 3 independent experiments