Figure 4
Figure 4. Kinetics of antigen-specific CD107a up-regulation. (A) Primary T cells transduced with the Tax-TCR and affinity-matured TCR and having undergone 3 rounds of weekly peptide stimulation had the following CD8+ and tetramer+ expression: Tax TCR = 92%; M1-TCR = 96%; M2-TCR = 95%; and M3-TCR = 98%. T cells (2 × 105) were stimulated with 2 × 105 T2 cells loaded with saturating concentration of pTax peptide in the presence of anti-CD107a, brefeldin A, and GolgiStop for the stated times. (A) Shown are the percentages of CD8+CD107a+ cells as a proportion of the tetramer-positive cells. The figure represents 4 individual experiments that demonstrated similar results. (B) Primary T cells transduced with the Tax-TCR and M3-TCR and having undergone 3 rounds of weekly peptide stimulation had the following percentage CD8+tetramer+ T cells: Tax TCR = 98%; M3-TCR = 99%. T cells (2 × 105) were stimulated with 2 × 105 T2 cells loaded with saturating concentration of pTax peptide or pHuD peptide in the presence of anti-CD107a, brefeldin A, and GolgiStop for the stated times. (B) Shown are the percentages of CD8+CD107a+ cells as a proportion of the tetramer-positive cells. Stimulation with T2 cells pulsed with the control pWT235 peptide or without peptide did not result in CD107 up-regulation. This figure represents 3 individual experiments that showed similar results. (C) Primary T cells transduced with the Tax-TCR and affinity-matured TCR and having undergone 5 rounds of weekly peptide stimulation had the following percentage CD8+tetramer+ T cells: Tax TCR = 91%; M1-TCR = 89%; M2-TCR = 86%; and M3-TCR = 86%. T cells (2 × 105) were stimulated with 2 × 105 cells loaded with decreasing concentrations of pTax peptide or pHuD peptide in the presence of anti-CD107a, brefeldin A, and Golgi Stop. CD107a expression was determined after a 5-hour stimulation period. Stimulation with T2 cells pulsed with the control pWT235 peptide or without peptide did not result in CD107 up-regulation. Shown is a representative of 3 individual experiments that showed similar results.

Kinetics of antigen-specific CD107a up-regulation. (A) Primary T cells transduced with the Tax-TCR and affinity-matured TCR and having undergone 3 rounds of weekly peptide stimulation had the following CD8+ and tetramer+ expression: Tax TCR = 92%; M1-TCR = 96%; M2-TCR = 95%; and M3-TCR = 98%. T cells (2 × 105) were stimulated with 2 × 105 T2 cells loaded with saturating concentration of pTax peptide in the presence of anti-CD107a, brefeldin A, and GolgiStop for the stated times. (A) Shown are the percentages of CD8+CD107a+ cells as a proportion of the tetramer-positive cells. The figure represents 4 individual experiments that demonstrated similar results. (B) Primary T cells transduced with the Tax-TCR and M3-TCR and having undergone 3 rounds of weekly peptide stimulation had the following percentage CD8+tetramer+ T cells: Tax TCR = 98%; M3-TCR = 99%. T cells (2 × 105) were stimulated with 2 × 105 T2 cells loaded with saturating concentration of pTax peptide or pHuD peptide in the presence of anti-CD107a, brefeldin A, and GolgiStop for the stated times. (B) Shown are the percentages of CD8+CD107a+ cells as a proportion of the tetramer-positive cells. Stimulation with T2 cells pulsed with the control pWT235 peptide or without peptide did not result in CD107 up-regulation. This figure represents 3 individual experiments that showed similar results. (C) Primary T cells transduced with the Tax-TCR and affinity-matured TCR and having undergone 5 rounds of weekly peptide stimulation had the following percentage CD8+tetramer+ T cells: Tax TCR = 91%; M1-TCR = 89%; M2-TCR = 86%; and M3-TCR = 86%. T cells (2 × 105) were stimulated with 2 × 105 cells loaded with decreasing concentrations of pTax peptide or pHuD peptide in the presence of anti-CD107a, brefeldin A, and Golgi Stop. CD107a expression was determined after a 5-hour stimulation period. Stimulation with T2 cells pulsed with the control pWT235 peptide or without peptide did not result in CD107 up-regulation. Shown is a representative of 3 individual experiments that showed similar results.

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