iMacs cultured in vitro retain the ability to suppress CTL generation.
Gr-1+ splenocytes from mice immunized with IL-2-rVV were enriched through panning with the specific mAb, and cultured in complete medium. After 6 days, adherent cells were collected in PBS-EDTA by gentle scraping, stained with different mAbs (A), and tested for their suppressive activity (B). (A) The percentage of adherent cells positive for any given marker is reported after subtraction of background staining with isotype-control mAbs. (B) Adherent cells were added at a final concentration of 3% of the total number of cells present in β-gal peptide-stimulated cultures of splenocytes from mice previously immunized with rAd-β-gal (immune). After a 5-day incubation, cultures were assayed in a51Cr-release assay against either CT26 cells (▪) or CT26 cells pulsed with the β-gal peptide (●). E:T cell ratios are indicated.
