Skip to main content
. 2017 Jun 20;9(8):1067–1087. doi: 10.15252/emmm.201707567

Figure EV3. Cytotoxicity and T‐cell activation by EnAd expressing EpCAM BiTE in DLD cells.

Figure EV3

  • A, B
    Cytotoxicity for infected DLD cells in the absence (A) or presence of T cells (B). DLD cells were infected and co‐cultured with T cells, and cytotoxicity was measured by LDH release at the specified time points.
  • C, D
    T cells from (B) were harvested and stained for activation markers CD69 (C) or CD25 (D) and analysed via flow cytometry.
  • E
    Quantification of EpCAM BiTE produced from DLD cells infected with recombinant viruses. Standard curve of LDH released (Abs) of DLD cells in co‐culture with CD3+ cells and serial dilutions of a known quantity of recombinant EpCAM BiTE (Ei). In parallel, co‐cultures were incubated with diluted supernatants (10,000‐fold) from 3‐day infected DLD cells (Eii). Standard curve allowed the approximate determination of EpCAM BiTE produced at 165 and 50 μg per million DLD cells for EnAd‐CMV‐EpCAMBiTE and EnAd‐SA‐EpCAMBiTE, respectively.
Data information: Each condition was measured in biological triplicate (A–D) or duplicate (E) and represented as mean ± SD (A–D). Significance was assessed by comparison to uninfected control wells using a one‐way ANOVA test with Tukey's post hoc analysis, **P < 0.01, ***P < 0.001.Source data are available online for this figure.