![Figure 2. Properties of T-cell responses induced by TGN1412 in high-density precultured PBMCs. (A) Precultured cells were stimulated with various mAb concentrations, and TNF was determined in supernatants after 24 hours. (B) Intracellular cytokine staining of high-density precultured PBMCs after stimulation with 1 μg/mL TGN1412 or OKT3. For optimal resolution, a responder with a particularly high TNF response was chosen. (C) Fresh and precultured PBMCs were prepared and cultured as described in legends to Figure 1A and B, respectively. Proliferation was determined by [3H]thymidine incorporation between days 2 and 3. (D) Proliferation of high-density precultured PBMCs by Ki67 staining, after 3-day stimulation with 1 μg/mL TGN1412 or OKT3. A different donor than in panel B is shown. (E) Fresh PBMCs were either stimulated immediately or placed in high density for 24 or 48 hours. They were then stimulated under standard conditions, and proliferation was determined by [3H]thymidine incorporation between days 2 and 3. (F) TNF release after TGN1412 stimulation of fresh and precultured PBMCs mixed at different ratios during the stimulation assay. Data are mean ± SD of triplicate samples.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/118/26/10.1182_blood-2010-12-319780/4/zh89991181560002.jpeg?Expires=1724226800&Signature=pjQgP1Mlpslp~NoTp-3Ks8l4AJ760xjVA9-TV74o-JltlM6hRAfKacPOPRzYROD1GUzF9lYTIE2RAAxghaUZDKhD5tlUo8fmYHGYkI49XwHJ8g5Irygfl83oCD4NKrh00QGUw~qj3~bGG66xDB3A7S0t3Lk3J2wYWpao6pZlFBK1ILeVCmkIbErjlRsX-qTzErDDDyNfxjEjKeXWeNash6cxEkNRzQeiHzx0eK8kHAgHUMFnpN9ByT2AJL2IrqzdGkqW5L79Q8ndm4SNNgKq37K5TSr4NxuPLz9BBZqZaNzoKCkiFx9TT12q7fiaOVUjlU5~WMR-UDrEehtSl-c5vQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Properties of T-cell responses induced by TGN1412 in high-density precultured PBMCs. (A) Precultured cells were stimulated with various mAb concentrations, and TNF was determined in supernatants after 24 hours. (B) Intracellular cytokine staining of high-density precultured PBMCs after stimulation with 1 μg/mL TGN1412 or OKT3. For optimal resolution, a responder with a particularly high TNF response was chosen. (C) Fresh and precultured PBMCs were prepared and cultured as described in legends to Figure 1A and B, respectively. Proliferation was determined by [3H]thymidine incorporation between days 2 and 3. (D) Proliferation of high-density precultured PBMCs by Ki67 staining, after 3-day stimulation with 1 μg/mL TGN1412 or OKT3. A different donor than in panel B is shown. (E) Fresh PBMCs were either stimulated immediately or placed in high density for 24 or 48 hours. They were then stimulated under standard conditions, and proliferation was determined by [3H]thymidine incorporation between days 2 and 3. (F) TNF release after TGN1412 stimulation of fresh and precultured PBMCs mixed at different ratios during the stimulation assay. Data are mean ± SD of triplicate samples.
