Figure 5.

Nef enhances activation in primary CD4 T cells. Purified CD4 T cells were transduced or not (NT) with the retroviral vectors as shown. Seven days after selection in G418, cells were incubated in the absence of exogenous stimulation overnight. Two hundred thousand rested cells and 6 × 10⁵ beads—precoated with anti-CD3 and anti-CD28 mAbs—were added to each well of a U-bottom, 96-well plate. Cells were incubated at 37°C, and supernatants were collected at various times to quantitate IL-2 secretion by standard ELISA methods. Determinations at 22 hr were averages from triplicate wells with a SD of 6.4% or less.