![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=1736047362&Signature=FhSYTFHL0qo83GFTbtVCgQThaw~Yg-bjjJitYiyfHdvTAHk7-4T~I-XRktAcHtjPZ1R25GciMkKfqwqVFaoPs-ywEo72NJXyU7H~64a6Y-fuHnV7DgoA~ZFD9p4sfPeCym~h5VT-bsdycO-VJ8-4yeocV25ctViUEwvHwTfmLxBzW15HtyGdU-hZ90aTsjx1GXbPP58pkdV9BcsBDzFG1bWXSYZ2Gp4otniwx68D21QHC0l2bhkScj8dsJO95zTdeb1MJ89Hv0DbzB6LR84hdMZ8saCQEiBv0Dhck048OT8kWBjILIndJic~nbFiycminQoXLCNEKNez6snfnA5zVA__&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.
