Summary of binding and cytotoxicity of ESK2 no. 34 on a panel of normal hematopoietic cells and nonhematopoietic cells
| Hematopoietic cells from healthy donors . | |||||
|---|---|---|---|---|---|
| Whole blood (binding/cytotoxicity) . | PBMCs (binding/cytotoxicity) . | ||||
| . | HLA-A2+ . | HLA-A2− . | . | HLA-A2+ . | HLA-A2− . |
| Neutrophils (CD15+) | Neg/neg | Neg/neg | T cells (CD3+) | Neg/neg | Neg/neg |
| Monocytes (CD33+) | Neg/neg | Neg/neg | Monocytes (CD33+) | Neg/neg | Neg/neg |
| Lymphocytes (CD45+) | 4%+/neg | Neg/neg | Monocytes (CD14+) | Neg/neg | Neg/neg |
| B cells (CD19+) | 2%-3%/neg | Neg/neg | |||
| Hematopoietic cells from healthy donors . | |||||
|---|---|---|---|---|---|
| Whole blood (binding/cytotoxicity) . | PBMCs (binding/cytotoxicity) . | ||||
| . | HLA-A2+ . | HLA-A2− . | . | HLA-A2+ . | HLA-A2− . |
| Neutrophils (CD15+) | Neg/neg | Neg/neg | T cells (CD3+) | Neg/neg | Neg/neg |
| Monocytes (CD33+) | Neg/neg | Neg/neg | Monocytes (CD33+) | Neg/neg | Neg/neg |
| Lymphocytes (CD45+) | 4%+/neg | Neg/neg | Monocytes (CD14+) | Neg/neg | Neg/neg |
| B cells (CD19+) | 2%-3%/neg | Neg/neg | |||
| Nonhematopoietic normal cells . | ||||
|---|---|---|---|---|
| . | WT1 mRNA expression . | HLA-A2 expression . | Binding . | Cytotoxicity . |
| Adrenal microvascular endothelial cells | +++ | Neg | Neg | Neg |
| Renal glomerular endothelial cells | +++ | Pos | Neg | Neg |
| Cardiac myocytes | NT | Neg | Neg | Neg |
| Testicular endothelial cells | NT | Pos | Neg | Neg |
| Nonhematopoietic normal cells . | ||||
|---|---|---|---|---|
| . | WT1 mRNA expression . | HLA-A2 expression . | Binding . | Cytotoxicity . |
| Adrenal microvascular endothelial cells | +++ | Neg | Neg | Neg |
| Renal glomerular endothelial cells | +++ | Pos | Neg | Neg |
| Cardiac myocytes | NT | Neg | Neg | Neg |
| Testicular endothelial cells | NT | Pos | Neg | Neg |
WT1 messenger RNA (mRNA) expression was measured by reverse transcription polymerase chain reaction, as previously described.16 Binding to normal cells was determined by ESK2 no. 34 mouse immunoglobulin G1 (3 μg/mL) over control isotype staining. Cytotoxicity was measured by ESK2 no. 34 BiTE at concentrations of 0.1, 1, or 10 μg/mL in standard 51Cr-release assay as described in “Materials and methods.”
For hematopoietic cells, the data are representative of 6 donors.
“+++” indicates 104 to 105 increases of WT1 transcript over negative control cells.
NT, not tested; Pos, positive (≥20-fold increase over isotype control staining for HLA-A2; BB7.2 clone); Neg, negative (no increase over isotype control staining).