Inhibition of cytotoxicity mediated by alloantigen-specific CD4+ and CD8+ CTLs by EGTA and CMA
| Donor . | Stimulator . | Phenotype . | Effector . | % Specific Lysis of Target Cells* . | |||||
|---|---|---|---|---|---|---|---|---|---|
| Allo LCL (Fas+/−) . | Allo LCL (Fas−/−) . | ||||||||
| None . | EGTA . | CMA . | None . | EGTA . | CMA . | ||||
| M.Y. | Fas+/− | CD4 | Bulk #1-1 | 45.8 | 0.8 | 1.5 | 43.0 | 0 | 0 |
| Clone #1-1 | 76.1 | 2.6 | 0.5 | 75.0 | 0.1 | 0.9 | |||
| Clone #1-2 | 52.8 | 0.3 | 0.1 | 49.8 | 0.1 | 0 | |||
| CD8 | Clone #2-1 | 78.0 | 2.0 | 0.4 | 76.9 | 2.0 | 1.7 | ||
| Fas−/− | CD4 | Bulk #3-1 | 34.0 | 0 | 0.3 | 38.9 | 0.7 | 1.0 | |
| Clone #3-1 | 69.2 | 0.1 | 0.5 | 65.8 | 0 | 0.8 | |||
| Clone #3-2 | 60.0 | 1.0 | 0 | 58.2 | 0 | 0 | |||
| CD8 | Clone #4-1 | 78.9 | 2.1 | 1.0 | 76.3 | 0.8 | 0.3 | ||
| H.O. | Fas+/− | CD4 | Bulk #5-1 | 42.9 | 0.5 | 1.6 | 40.5 | 0.6 | 0 |
| Clone #5-1 | 65.2 | 0.6 | 1.0 | 66.6 | 0 | 1.7 | |||
| Clone #5-2 | 54.2 | 0 | 0 | 51.0 | 0.4 | 0 | |||
| CD8 | Clone #6-1 | 65.0 | 0.9 | 1.5 | 56.9 | 0.8 | 0 | ||
| Fas−/− | CD4 | Bulk #7-1 | 46.9 | 0.8 | 0 | 45.9 | 0.3 | 0 | |
| Clone #7-1 | 67.7 | 2.0 | 1.6 | 69.8 | 1.1 | 1.3 | |||
| Clone #7-2 | 56.9 | 0.5 | 0.8 | 54.9 | 0.3 | 1.8 | |||
| CD8 | Clone #8-1 | 75.2 | 2.9 | 0.7 | 70.8 | 0.8 | 0 | ||
| Donor . | Stimulator . | Phenotype . | Effector . | % Specific Lysis of Target Cells* . | |||||
|---|---|---|---|---|---|---|---|---|---|
| Allo LCL (Fas+/−) . | Allo LCL (Fas−/−) . | ||||||||
| None . | EGTA . | CMA . | None . | EGTA . | CMA . | ||||
| M.Y. | Fas+/− | CD4 | Bulk #1-1 | 45.8 | 0.8 | 1.5 | 43.0 | 0 | 0 |
| Clone #1-1 | 76.1 | 2.6 | 0.5 | 75.0 | 0.1 | 0.9 | |||
| Clone #1-2 | 52.8 | 0.3 | 0.1 | 49.8 | 0.1 | 0 | |||
| CD8 | Clone #2-1 | 78.0 | 2.0 | 0.4 | 76.9 | 2.0 | 1.7 | ||
| Fas−/− | CD4 | Bulk #3-1 | 34.0 | 0 | 0.3 | 38.9 | 0.7 | 1.0 | |
| Clone #3-1 | 69.2 | 0.1 | 0.5 | 65.8 | 0 | 0.8 | |||
| Clone #3-2 | 60.0 | 1.0 | 0 | 58.2 | 0 | 0 | |||
| CD8 | Clone #4-1 | 78.9 | 2.1 | 1.0 | 76.3 | 0.8 | 0.3 | ||
| H.O. | Fas+/− | CD4 | Bulk #5-1 | 42.9 | 0.5 | 1.6 | 40.5 | 0.6 | 0 |
| Clone #5-1 | 65.2 | 0.6 | 1.0 | 66.6 | 0 | 1.7 | |||
| Clone #5-2 | 54.2 | 0 | 0 | 51.0 | 0.4 | 0 | |||
| CD8 | Clone #6-1 | 65.0 | 0.9 | 1.5 | 56.9 | 0.8 | 0 | ||
| Fas−/− | CD4 | Bulk #7-1 | 46.9 | 0.8 | 0 | 45.9 | 0.3 | 0 | |
| Clone #7-1 | 67.7 | 2.0 | 1.6 | 69.8 | 1.1 | 1.3 | |||
| Clone #7-2 | 56.9 | 0.5 | 0.8 | 54.9 | 0.3 | 1.8 | |||
| CD8 | Clone #8-1 | 75.2 | 2.9 | 0.7 | 70.8 | 0.8 | 0 | ||
Cytotoxicity of CD4+ and CD8+ CTL bulk lines and clones generated by stimulation of PBMCs isolated from unrelated donors with HLA class I and class II-mismatched Fas+/− or Fas−/− LCL against Fas+/− and Fas−/− target cells in the presence or absence of EGTA or CMA, as detailed in “Materials and methods.” Effector/target ratios were 5:1.