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. 2014 Jul 23;5(15):6466–6483. doi: 10.18632/oncotarget.2238

Figure 4. Specific redirected lysis of malignant target cell lines mediated by the 19-3 BiTE and triplebody 19-3-19 in conjunction with effector T cells.

Figure 4

Standard Calcein Release Assays with an E : T ratio of 10 : 1 (MNCs : target cells) and a duration of 3 hrs were performed unless indicated otherwise. Error bars indicate SEM. (A) Dose-Response-Curve for SEM cells (triangles: triplebody 19-3-19; squares: 19-3 BiTE; cross: specificity control Her2-3-Her2; n=4). (B) Maximum specific lysis of several CD19-positive malignant B-lymphoid cell lines (n=4 each) induced by 19-3 (white bars), triplebody 19-3-19 (black bars) and specificity control Her2-3-Her2 (grey bars), respectively. (C) Serial lysis of unlabeled SEM target cells mediated by triplebody 19-3-19 at different E : T ratios. The reaction mixture (purified T cells and SEM target cells in RPMI1640 GlutaMAX medium containing 10% FCS) was incubated with or without 1nM triplebody overnight (approx. 15 hrs) (n = 3). The total number of viable cells in each reaction was determined and the fraction of CD19-positive target cells was established by FACS analysis.