Lack of expression of the exogenous gene in granulocytes and macropahges. (A) Southern blotting of genomic DNA prepared from PEC 24 and 96 hours after thioglycollate treatment and spleen from two representative mice reconstituted 6 months earlier with 20 × 106 cells of T-cell–depleted spleen cells infected with pAsADA. DNA was digested with SacI and probed with aneo-specific probe. SacI-digested pAsADA plasmid DNA equivalent to 0.25 and 1 copies/cell are as indicated. (B) Human and mouse ADA protein assay. Cell lysates prepared from 1 × 106 cells from the same spleen and PEC samples used for Southern blotting were electrophoresed on a cellulose acetate plate to separate human from murine ADA enzymes. ADA activity was detected by the standard colorimetric enzymatic assay. PEC were analyzed 24 hours (granulocytes) and 96 hours (macrophages) after thioglycollate induction, respectively.
