Anti-Human CD3 Recombinant Antibody (OKT3) (CAT#: TAB-0416CL)

Figure 1 Soluble anti-CD3 mAbs induce death of preactivated and cycling human peripheral T cells.

T cells were preactivated for 24 h (left panel) or 72 h (right panel) by solid phase-bound anti-CD3 mAb, BC3. T cells were harvested, one aliquot was used for cell cycle analysis, and other aliquots were exposed for 24 h to 0.2 μg/ml of the anti-CD3 mAbs, HuM291-IgG2M3 or OKT3, or to a murine IgG2a mAb of irrelevant specificity in the presence of 50 IU/ml IL-2 in CM containing 10% normal human serum.

Carpenter, P. A., Pavlovic, S., Tso, J. Y., Press, O. W., Gooley, T., Yu, X. Z., & Anasetti, C. (2000). Non-Fc receptor-binding humanized anti-CD3 antibodies induce apoptosis of activated human T cells. The Journal of Immunology, 165(11), 6205-6213.

Figure 2 HuM291 induces greater and more sustained ERK phosphorylation in activated human T cells than OKT3.

Cells were cultured in medium alone or were stimulated at 37°C with 50 μg/μl HuM291 or 50 μg/μl OKT3 for the indicated time in minutes. Activated cells stimulated with PMA (15 ng/ml) plus ionomycin (700 ng/ml) for 10 min were analyzed as a positive control for ERK phosphorylation. Cells receiving 50 μg/μl anti-human IgG2 or 50 μg/μl anti-mouse IgG2a for 10 min were analyzed as additional negative controls for ERK phosphorylation. Whole-cell lysates were prepared, and 20 μg of protein was separated by SDS-PAGE as described in Materials and Methods. ERK phosphorylation (p-ERK1 and p-ERK2) was determined by Western blotting with an anti-phospho-ERK mAb (top panel). Subsequently, the same nitrocellulose membrane was stripped and reprobed with an anti-ERK2 mAb to determine ERK2 expression for each time interval and condition (bottom panel). A representative experiment of three performed is shown.

Carpenter, P. A., Pavlovic, S., Tso, J. Y., Press, O. W., Gooley, T., Yu, X. Z., & Anasetti, C. (2000). Non-Fc receptor-binding humanized anti-CD3 antibodies induce apoptosis of activated human T cells. The Journal of Immunology, 165(11), 6205-6213.

Figure 3 Anti-CD3 mAb variants induce more T cell death and release of IFN-γ than do murine anti-CD3 mAbs OKT3 or M291.

Supernatants from samples cultured in parallel to those for the cell death assays, but without exogenous IL-2, and with CM containing FCS instead of pooled human serum were assayed for the cytokines indicated.

Carpenter, P. A., Pavlovic, S., Tso, J. Y., Press, O. W., Gooley, T., Yu, X. Z., & Anasetti, C. (2000). Non-Fc receptor-binding humanized anti-CD3 antibodies induce apoptosis of activated human T cells. The Journal of Immunology, 165(11), 6205-6213.

Figure 1 Anti-CD3 Antibody OKT3 (TAB-0416CL) Induced PBMCs Proliferation

Peripheral blood mononuclear cells (PBMCs) were cultured and stimulated with the anti-CD3 antibody OKT3 (Cat# TAB-0416CL, Creative biolabs) at a series of concentrations (0 μg/mL, 0.01 μg/mL, 0.05 μg/mL, 1 μg/mL, and 5 μg/mL).