Results of CAF activity in subcellular fractions of CD8+ cells
Subject . | CAF activity, % suppression* . | Fraction . | Granzyme B concentration, nM . | Total volume, μL . | Total granzyme B yield, ng . | CAF activity† . |
|---|---|---|---|---|---|---|
| A | 46 | Cytoplasm, organelle free | 0 | 2000 | 0 | + |
| Soluble granule contents | 158 | 1500 | 7580 | − | ||
| Granule membrane-associated | 448 | 200 | 2870 | − | ||
| B | 61 | Cytoplasm, organelle free | 1 | 1000 | 30 | + |
| Soluble granule contents | 62 | 1000 | 1973 | − | ||
| Granule membrane-associated | 193 | 100 | 617 | − | ||
| C | 7 | Cytoplasm, organelle free | 25 | 1000 | 792 | − |
| Soluble granule contents | 154 | 1000 | 4937 | − | ||
| Granule membrane-associated | 1514 | 100 | 4845 | − |
Subject . | CAF activity, % suppression* . | Fraction . | Granzyme B concentration, nM . | Total volume, μL . | Total granzyme B yield, ng . | CAF activity† . |
|---|---|---|---|---|---|---|
| A | 46 | Cytoplasm, organelle free | 0 | 2000 | 0 | + |
| Soluble granule contents | 158 | 1500 | 7580 | − | ||
| Granule membrane-associated | 448 | 200 | 2870 | − | ||
| B | 61 | Cytoplasm, organelle free | 1 | 1000 | 30 | + |
| Soluble granule contents | 62 | 1000 | 1973 | − | ||
| Granule membrane-associated | 193 | 100 | 617 | − | ||
| C | 7 | Cytoplasm, organelle free | 25 | 1000 | 792 | − |
| Soluble granule contents | 154 | 1000 | 4937 | − | ||
| Granule membrane-associated | 1514 | 100 | 4845 | − |
Results are representative of 6 subjects studied.
Granzyme B concentration was measured using an enzymatic assay (“Materials and methods”).
Total granzyme B yield was calculated from the respective concentration and total volume.
Total volume of each fraction was approximately 170- to 300-fold (1700- to 3000-fold in membrane-associated fraction) less per number of cells than that used to obtain the CAF-active culture supernatants.
See also Figure 2 regarding CAF activity in the subcellular fractions.
Activity of CAF was measured as described in “Materials and methods.” Fluids were collected at 9 days (subject A) and 7 days (subjects B and C) after CD8+ cell stimulation. 11 Total number of CD8+ cells processed was 13.7 × 108 (subject A) and 5.0 × 108 (subjects B and C).
Activity measured in subcellular fractions.