Figure 4.
Figure 4. Inhibition of TPT on the allostimulatory activity of DCs and LPS-stimulated DCs. (A) Human monocytes were cultured with 50 ng/mL GM-CSF and 50 ng/mL IL-4 in the presence of TPT (TPT[D2-7], 2.5 nM) or medium alone (control). After 7 days, DCs were extensively washed, irradiated (30 Gy [3000 rad]), and added at different concentrations to 1 × 105/well purified allogeneic responder T cells in a 96-well flat-bottom plates. (B) In a second type of experiment, TPT2-treated DCs (TPT[D2-7], 2.5 nM) and control DCs were washed and cultured for another 48 hours with LPS (10 ng/mL). Aliquots of control DCs were cultured with LPS in the presence of TPT (TPT[D8-9], 2.5 nM). After 48 hours, DCs were thoroughly washed and irradiated (30 Gy [3000 rad]), and added in graded doses to 1 × 105/well purified allogeneic responder T cells in a 96-well flat-bottom plates. Each group was tested in triplicate, and thymidine incorporation was measured on day 5 by a 15-hour pulse with [3H] thymidine. The data shown are representative of 3 independent experiments with similar results. Error bars indicate SD.

Inhibition of TPT on the allostimulatory activity of DCs and LPS-stimulated DCs. (A) Human monocytes were cultured with 50 ng/mL GM-CSF and 50 ng/mL IL-4 in the presence of TPT (TPT[D2-7], 2.5 nM) or medium alone (control). After 7 days, DCs were extensively washed, irradiated (30 Gy [3000 rad]), and added at different concentrations to 1 × 105/well purified allogeneic responder T cells in a 96-well flat-bottom plates. (B) In a second type of experiment, TPT2-treated DCs (TPT[D2-7], 2.5 nM) and control DCs were washed and cultured for another 48 hours with LPS (10 ng/mL). Aliquots of control DCs were cultured with LPS in the presence of TPT (TPT[D8-9], 2.5 nM). After 48 hours, DCs were thoroughly washed and irradiated (30 Gy [3000 rad]), and added in graded doses to 1 × 105/well purified allogeneic responder T cells in a 96-well flat-bottom plates. Each group was tested in triplicate, and thymidine incorporation was measured on day 5 by a 15-hour pulse with [3H] thymidine. The data shown are representative of 3 independent experiments with similar results. Error bars indicate SD.

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