Summary of CTLA-4 expression in human normal hematopoietic cells
| Hematopoietic normal cells . | Surface . | Cytoplasm . | ||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| scFv no. . | mAbs . | scFv no. . | mAbs . | |||||||||||||
| 3 . | 40 . | 55 . | 67 . | 83 . | 26 . | BN13 . | IgG1 . | 3 . | 40 . | 55 . | 67 . | 83 . | 26 . | BN13 . | IgG1 . | |
| PBMCs | − | − | − | − | − | − | − | − | ++ | ++ | ++ | ++ | ++ | − | ++ | − |
| PBMCs plus PMA/PHA | + | ++ | ++ | ++ | ++ | − | + | − | ++ | ++ | ++ | ++ | ++ | − | ++ | − |
| T cells | − | − | − | − | − | − | − | − | ++ | ++ | ++ | ++ | ++ | − | ++ | − |
| T cells plus PMA/PHA | + | + | ++ | ++ | ++ | − | + | − | ++ | ++ | ++ | ++ | ++ | − | ++ | − |
| B cells | − | − | − | − | − | − | − | − | + | ++ | + | ++ | ++ | − | ++ | − |
| B cells plus PMA | ± | + | + | + | + | − | + | − | + | ++ | ++ | ++ | ++ | − | ++ | − |
| Granulocytes | − | − | − | − | − | − | − | − | − | − | − | − | − | − | − | − |
| Granulocytes plus G-CSF | + | + | + | + | + | − | + | − | + | + | + | + | + | − | + | − |
| Stem cells (CD34+) | − | − | − | − | − | − | − | − | + | ++ | ++ | + | + | − | ++ | − |
| Stem cells (CD34+) plus GM-CSF | + | + | + | + | ++ | − | ++ | − | + | ++ | ++ | ++ | ++ | − | ++ | − |
| Monocytes (CD14+) | − | − | ± | ± | ± | − | ± | − | + | + | + | + | + | − | + | − |
| Monocytes (CD14+) plus GM-CSF | ++ | ++ | ++ | ++ | ++ | − | ++ | − | ++ | ++ | ++ | ++ | ++ | − | ++ | − |
| Hematopoietic normal cells . | Surface . | Cytoplasm . | ||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| scFv no. . | mAbs . | scFv no. . | mAbs . | |||||||||||||
| 3 . | 40 . | 55 . | 67 . | 83 . | 26 . | BN13 . | IgG1 . | 3 . | 40 . | 55 . | 67 . | 83 . | 26 . | BN13 . | IgG1 . | |
| PBMCs | − | − | − | − | − | − | − | − | ++ | ++ | ++ | ++ | ++ | − | ++ | − |
| PBMCs plus PMA/PHA | + | ++ | ++ | ++ | ++ | − | + | − | ++ | ++ | ++ | ++ | ++ | − | ++ | − |
| T cells | − | − | − | − | − | − | − | − | ++ | ++ | ++ | ++ | ++ | − | ++ | − |
| T cells plus PMA/PHA | + | + | ++ | ++ | ++ | − | + | − | ++ | ++ | ++ | ++ | ++ | − | ++ | − |
| B cells | − | − | − | − | − | − | − | − | + | ++ | + | ++ | ++ | − | ++ | − |
| B cells plus PMA | ± | + | + | + | + | − | + | − | + | ++ | ++ | ++ | ++ | − | ++ | − |
| Granulocytes | − | − | − | − | − | − | − | − | − | − | − | − | − | − | − | − |
| Granulocytes plus G-CSF | + | + | + | + | + | − | + | − | + | + | + | + | + | − | + | − |
| Stem cells (CD34+) | − | − | − | − | − | − | − | − | + | ++ | ++ | + | + | − | ++ | − |
| Stem cells (CD34+) plus GM-CSF | + | + | + | + | ++ | − | ++ | − | + | ++ | ++ | ++ | ++ | − | ++ | − |
| Monocytes (CD14+) | − | − | ± | ± | ± | − | ± | − | + | + | + | + | + | − | + | − |
| Monocytes (CD14+) plus GM-CSF | ++ | ++ | ++ | ++ | ++ | − | ++ | − | ++ | ++ | ++ | ++ | ++ | − | ++ | − |
Reactivity of different peripheral blood cells with a panel of FITC-conjugated anti-CTLA-4 scFvs and the commercial BN13 mAb by flow cytometry. Fluorescence intensity was scored as follows: −, negative; ±, weak; and + to ++, positive, with grading from 1 log to more than 1 log of histogram shift relative to the negative control. The FITC-conjugated anti-BSA scFv 26 and a FITC-conjugated mouse IgG1 mAb were used as negative controls for anti-CTLA-4 scFvs and BN13 mAb, respectively. All cell samples were tested either resting or in vitro activated, except granulocytes that were derived from in vivo G-CSF–treated donors, as described in “Patients, materials and methods.”