T-cell subset expansion and cell size after chemotherapy exposure (long-term)
| . | Control . | Dox . | 4HPCP . | Cytarabine . |
|---|---|---|---|---|
| Expansion (SD), fold | ||||
| Unstimulated | ||||
| Bulk | 0.71 (.22) | 0.52 (.26) | 0.42 (.10) | 0.50 (.10) |
| Naive | 0.99 (.13) | 0.82 (.20) | 0.41 (.11) | 0.60 (.10) |
| SCM | 1.29 (.11) | 1.10 (.24) | 0.53 (.10) | 0.73 (.11) |
| CM | 0.95 (.08) | 0.69 (.26) | 0.49 (.10) | 0.65 (.12) |
| EM | 0.76 (.24) | 0.67 (.19) | 0.82 (.15) | 0.66 (.10) |
| Stimulated | ||||
| Bulk | 4.10 (2.1) | 0.48 (.23) | 3.80 (.36) | 0.23 (.09) |
| Naive | 4.70 (1.4) | 0.75 (.24) | 0.89 (.12) | 0.33 (.10) |
| SCM | 4.00 (1.8) | 1.30 (.33) | 3.10 (.25) | 0.34 (.10) |
| CM | 5.70 (2.1) | 0.37 (.10) | 0.95 (.11) | 0.32 (.08) |
| EM | 2.10 (1.3) | 0.33 (.11) | 0.55 (.09) | 0.40 (.10) |
| Cell size (SD), fL | ||||
| Unstimulated | ||||
| Bulk | 186 (19) | 186 (20) | 182 (18) | 192 (18) |
| Naive | 186 (19) | 186 (21) | 189 (21) | 186 (18) |
| SCM | 186 (19) | 186 (20) | 190 (20) | 189 (18) |
| CM | 202 (21) | 204 (22) | 204 (20) | 203 (19) |
| EM | 203 (19) | 191 (20) | 208 (20) | 203 (19) |
| Stimulated | ||||
| Bulk | 405 (54) | 285 (26)* | 375 (36) | 208 (18)* |
| Naive | 460 (69) | 308 (42)* | 297 (37)* | 226 (19)* |
| SCM | 505 (59) | 365 (58) | 461 (59) | 189 (18)* |
| CM | 391 (49) | 263 (41)* | 230 (29)* | 203 (19)* |
| EM | 294 (31) | 245 (24) | 229 (24) | 229 (20) |
| . | Control . | Dox . | 4HPCP . | Cytarabine . |
|---|---|---|---|---|
| Expansion (SD), fold | ||||
| Unstimulated | ||||
| Bulk | 0.71 (.22) | 0.52 (.26) | 0.42 (.10) | 0.50 (.10) |
| Naive | 0.99 (.13) | 0.82 (.20) | 0.41 (.11) | 0.60 (.10) |
| SCM | 1.29 (.11) | 1.10 (.24) | 0.53 (.10) | 0.73 (.11) |
| CM | 0.95 (.08) | 0.69 (.26) | 0.49 (.10) | 0.65 (.12) |
| EM | 0.76 (.24) | 0.67 (.19) | 0.82 (.15) | 0.66 (.10) |
| Stimulated | ||||
| Bulk | 4.10 (2.1) | 0.48 (.23) | 3.80 (.36) | 0.23 (.09) |
| Naive | 4.70 (1.4) | 0.75 (.24) | 0.89 (.12) | 0.33 (.10) |
| SCM | 4.00 (1.8) | 1.30 (.33) | 3.10 (.25) | 0.34 (.10) |
| CM | 5.70 (2.1) | 0.37 (.10) | 0.95 (.11) | 0.32 (.08) |
| EM | 2.10 (1.3) | 0.33 (.11) | 0.55 (.09) | 0.40 (.10) |
| Cell size (SD), fL | ||||
| Unstimulated | ||||
| Bulk | 186 (19) | 186 (20) | 182 (18) | 192 (18) |
| Naive | 186 (19) | 186 (21) | 189 (21) | 186 (18) |
| SCM | 186 (19) | 186 (20) | 190 (20) | 189 (18) |
| CM | 202 (21) | 204 (22) | 204 (20) | 203 (19) |
| EM | 203 (19) | 191 (20) | 208 (20) | 203 (19) |
| Stimulated | ||||
| Bulk | 405 (54) | 285 (26)* | 375 (36) | 208 (18)* |
| Naive | 460 (69) | 308 (42)* | 297 (37)* | 226 (19)* |
| SCM | 505 (59) | 365 (58) | 461 (59) | 189 (18)* |
| CM | 391 (49) | 263 (41)* | 230 (29)* | 203 (19)* |
| EM | 294 (31) | 245 (24) | 229 (24) | 229 (20) |
T cells from normal donors were sorted by flow cytometry and exposed to chemotherapy for 24 hours (doxorubicin at 400 nM, 4HPCP at 2.5 μM, and cytarabine at 3 mM), allowed to recover and then exposed to CD3/28 beads as in CAR manufacture (∼13 days total). Chemotherapy almost uniformly blocked expansion of all T-cell subsets with the exception of 4HPCP (cyclophosphamide)-exposed SCM cells. When looking at the ability of the cells to blast, doxorubicin reduced blasting in most subsets, 4HPCP blocked naive and CM blasting, and cytarabine blocked essentially all cells. This suggests that the action of chemotherapy on each subset is different. Bold values represent statistically significant changes to control (P < .05).
Dox, doxorubicin; SD, standard deviation.
P < .05.