FIG. 4.
Stability of the complexes formed between H-2Db and the LCMV epitopes. RMA-S previously incubated at 26°C were pulsed with 1 μM peptide for 4 h at 37°C. After three washes at 4°C in phosphate-buffered saline containing 2% fetal calf serum, cells were resuspended in medium without fetal calf serum and then transferred to 37°C. At different times (0, 2, and 4 h), aliquots (2.5 × 105 cells) were removed and stained for H-2Db expression using the 28-14-8S mouse monoclonal antibody. Cells were then stained with a fluorescent (fluorescein isothiocyanate) secondary anti-mouse antibody and analyzed by flow cytometry. Results are expressed as percent fluorescence (100% at 0 h).