Fig. 5.
Neutralization of supernatants of cultures of clones 65 and 67 with anti-TNFβ MoAbs. Supernatants of 4-day cultures of clones 65 and 67 after stimulation of 30 Gy irradiated allogeneic PBLs having DRB1*1502 in the presence of 20 U/mL rIL-2 (initial cell concentration of 2 × 105 cells/mL), which exhibited cytotoxic activity against L929 cells as shown in Fig 4, were incubated with a saturating concentration of anti-TNFβ MoAb for 60 minutes at 37°C. The treated supernatants or nontreated controls were added to monolayers of L929 cells in the presence of 1 μg/mL actinomycin D. After an 18-hour incubation, the viable cells were stained with methylene blue and absorbance at 660 nm was measured.
