In vitro responses by mixed lymphocyte reaction (MLR) assay
| . | Early harvest (D2 or D3) DD SI . | Standard harvest (D5) DD SI . | D2 or D3 SI ratio to before treatment . | SI ratio (D2 or D3/D5) . | Antibody response (titer) . | Cytotoxic antibody . |
|---|---|---|---|---|---|---|
| Animal 16707/PBMCs* | ||||||
| Before treatment | 4.7 | 19 | 1.000 | 0.247 | — | — |
| D 15 | 8.1 | 24.1 | 1.723 | 0.336 | — | — |
| D 30 | 8.1 | 26.1 | 1.723 | 0.310 | Yes (1:2560) | Yes |
| Animal 16792/PBMCs* | ||||||
| Before treatment‡ | N/A | N/A | N/A | N/A | — | — |
| D 15 | 13 | 79 | N/A | 0.165 | — | — |
| D 30 | 2.7 | 12.8 | N/A | 0.211 | Yes (1:2560) | Yes |
| Animal 16649/UTCs IV* | ||||||
| Before treatment | 1.4 | 8.3 | 1.000 | 0.169 | — | — |
| D 15 | 1.3 | 6.1 | 0.929 | 0.213 | — | — |
| D 30 | 1.4 | 16.6 | 1.000 | 0.084 | — | — |
| D 60 | 1.5 | 17.4 | 1.071 | 0.086 | No | No |
| Animal 16481/UTCs IV* | ||||||
| Before treatment | 0.8 | 11 | 1.000 | 0.073 | — | — |
| D 15§ | N/A | N/A | N/A | N/A | — | — |
| D 30 | 1 | 5.1 | 1.250 | 0.196 | — | — |
| D 60 | 1.6 | 7.4 | 2.000 | 0.216 | No | No |
| Animal 16650/UTCs SC* | ||||||
| Before treatment | 3.1 | 53.7 | 1.000 | 0.058 | — | — |
| D 15 | 2.5 | 7.5 | 0.806 | 0.333 | — | — |
| D 30 | 5.9 | 15.2 | 1.903 | 0.388 | No | No |
| Animal 16633/UTCs SC* | ||||||
| Before treatment‡ | N/A | N/A | N/A | N/A | — | — |
| D 15 | 5.4 | 6.8 | N/A | 0.794 | — | — |
| D 30 | 2.6 | 4.6 | N/A | 0.565 | No | No |
| Animal 16843/CFA/UTCs* | ||||||
| Before treatment | 6.9 | 123 | 1.000 | 0.056 | — | — |
| D 15 | 4.1 | 4.2 | 0.594 | 0.976 | — | — |
| D 30 | 3.3 | 7.6 | 0.478 | 0.434 | — | — |
| After skin graft | 11.4 | 8.6 | 1.652 | 1.326 | Yes (1:640) | Yes |
| Animal 16844/CFA/UTCs* | ||||||
| Before treatment | 5.4 | 37.7 | 1.000 | 0.143 | — | — |
| D 15 | 3.2 | 5.6 | 0.593 | 0.571 | — | — |
| D 30 | 3.9 | 6.1 | 0.722 | 0.639 | — | — |
| After skin graft | 13.7 | 17 | 2.537 | 0.806 | Yes (1:640) | Yes |
| Animal 16854/aUTCs† | ||||||
| Before treatment | N/A | 221 | N/A | N/A | — | — |
| D 15 | 3.7 | 248 | N/A | 0.015 | — | — |
| D 30 | 0.44 | 1.3 | N/A | 0.338 | — | — |
| D 60 | 9.5 | 409 | N/A | 0.023 | — | — |
| After skin graft | 19 | 535 | N/A | 0.036 | Yes (1:640) | Yes |
| Animal 16922/aUTCs† | ||||||
| Before treatment | N/A | 31 | N/A | N/A | — | — |
| D 15 | 12 | 31 | N/A | 0.387 | — | — |
| D 30§ | N/A | N/A | N/A | N/A | — | — |
| D 60 | 3.4 | 489 | N/A | 0.007 | — | — |
| After skin graft | 20 | 186 | N/A | 0.108 | Yes (1:640) | Yes |
| Animal 17025/UTCs SC×3† | ||||||
| Before treatment | 4.2 | 51 | 1.000 | 0.082 | — | — |
| D 15 | 6.8 | 208 | 1.619 | 0.033 | — | — |
| D 45 | 19 | 426 | 4.524 | 0.045 | — | — |
| D 75 | 56 | 87 | 13.333 | 0.644 | Yes (after 2nd inj, >1:10240) | Yes (after 2nd inj) |
| After skin graft | 90 | 27 | 21.429 | 3.333 | — | — |
| Animal 17026/UTCs SC×3† | ||||||
| 2 Before treatment | 4.6 | 165 | 1.000 | 0.028 | — | — |
| D 15 | 8.2 | 223 | 1.783 | 0.037 | — | — |
| D 45 | 30 | 125 | 6.522 | 0.240 | — | — |
| D 75 | 29 | 0.58 | 6.304 | 50.000 | — | — |
| D 96 | 487 | 39 | 105.870 | 12.487 | — | — |
| After skin graft | 21 | 2.2 | 4.565 | 9.545 | Yes (after 2nd inj, 1:640) | Yes (after 2nd inj) |
| After skin graft repeat | 166 | 1.1 | 36.087 | 150.909 | — | — |
| . | Early harvest (D2 or D3) DD SI . | Standard harvest (D5) DD SI . | D2 or D3 SI ratio to before treatment . | SI ratio (D2 or D3/D5) . | Antibody response (titer) . | Cytotoxic antibody . |
|---|---|---|---|---|---|---|
| Animal 16707/PBMCs* | ||||||
| Before treatment | 4.7 | 19 | 1.000 | 0.247 | — | — |
| D 15 | 8.1 | 24.1 | 1.723 | 0.336 | — | — |
| D 30 | 8.1 | 26.1 | 1.723 | 0.310 | Yes (1:2560) | Yes |
| Animal 16792/PBMCs* | ||||||
| Before treatment‡ | N/A | N/A | N/A | N/A | — | — |
| D 15 | 13 | 79 | N/A | 0.165 | — | — |
| D 30 | 2.7 | 12.8 | N/A | 0.211 | Yes (1:2560) | Yes |
| Animal 16649/UTCs IV* | ||||||
| Before treatment | 1.4 | 8.3 | 1.000 | 0.169 | — | — |
| D 15 | 1.3 | 6.1 | 0.929 | 0.213 | — | — |
| D 30 | 1.4 | 16.6 | 1.000 | 0.084 | — | — |
| D 60 | 1.5 | 17.4 | 1.071 | 0.086 | No | No |
| Animal 16481/UTCs IV* | ||||||
| Before treatment | 0.8 | 11 | 1.000 | 0.073 | — | — |
| D 15§ | N/A | N/A | N/A | N/A | — | — |
| D 30 | 1 | 5.1 | 1.250 | 0.196 | — | — |
| D 60 | 1.6 | 7.4 | 2.000 | 0.216 | No | No |
| Animal 16650/UTCs SC* | ||||||
| Before treatment | 3.1 | 53.7 | 1.000 | 0.058 | — | — |
| D 15 | 2.5 | 7.5 | 0.806 | 0.333 | — | — |
| D 30 | 5.9 | 15.2 | 1.903 | 0.388 | No | No |
| Animal 16633/UTCs SC* | ||||||
| Before treatment‡ | N/A | N/A | N/A | N/A | — | — |
| D 15 | 5.4 | 6.8 | N/A | 0.794 | — | — |
| D 30 | 2.6 | 4.6 | N/A | 0.565 | No | No |
| Animal 16843/CFA/UTCs* | ||||||
| Before treatment | 6.9 | 123 | 1.000 | 0.056 | — | — |
| D 15 | 4.1 | 4.2 | 0.594 | 0.976 | — | — |
| D 30 | 3.3 | 7.6 | 0.478 | 0.434 | — | — |
| After skin graft | 11.4 | 8.6 | 1.652 | 1.326 | Yes (1:640) | Yes |
| Animal 16844/CFA/UTCs* | ||||||
| Before treatment | 5.4 | 37.7 | 1.000 | 0.143 | — | — |
| D 15 | 3.2 | 5.6 | 0.593 | 0.571 | — | — |
| D 30 | 3.9 | 6.1 | 0.722 | 0.639 | — | — |
| After skin graft | 13.7 | 17 | 2.537 | 0.806 | Yes (1:640) | Yes |
| Animal 16854/aUTCs† | ||||||
| Before treatment | N/A | 221 | N/A | N/A | — | — |
| D 15 | 3.7 | 248 | N/A | 0.015 | — | — |
| D 30 | 0.44 | 1.3 | N/A | 0.338 | — | — |
| D 60 | 9.5 | 409 | N/A | 0.023 | — | — |
| After skin graft | 19 | 535 | N/A | 0.036 | Yes (1:640) | Yes |
| Animal 16922/aUTCs† | ||||||
| Before treatment | N/A | 31 | N/A | N/A | — | — |
| D 15 | 12 | 31 | N/A | 0.387 | — | — |
| D 30§ | N/A | N/A | N/A | N/A | — | — |
| D 60 | 3.4 | 489 | N/A | 0.007 | — | — |
| After skin graft | 20 | 186 | N/A | 0.108 | Yes (1:640) | Yes |
| Animal 17025/UTCs SC×3† | ||||||
| Before treatment | 4.2 | 51 | 1.000 | 0.082 | — | — |
| D 15 | 6.8 | 208 | 1.619 | 0.033 | — | — |
| D 45 | 19 | 426 | 4.524 | 0.045 | — | — |
| D 75 | 56 | 87 | 13.333 | 0.644 | Yes (after 2nd inj, >1:10240) | Yes (after 2nd inj) |
| After skin graft | 90 | 27 | 21.429 | 3.333 | — | — |
| Animal 17026/UTCs SC×3† | ||||||
| 2 Before treatment | 4.6 | 165 | 1.000 | 0.028 | — | — |
| D 15 | 8.2 | 223 | 1.783 | 0.037 | — | — |
| D 45 | 30 | 125 | 6.522 | 0.240 | — | — |
| D 75 | 29 | 0.58 | 6.304 | 50.000 | — | — |
| D 96 | 487 | 39 | 105.870 | 12.487 | — | — |
| After skin graft | 21 | 2.2 | 4.565 | 9.545 | Yes (after 2nd inj, 1:640) | Yes (after 2nd inj) |
| After skin graft repeat | 166 | 1.1 | 36.087 | 150.909 | — | — |
D 2 or D 3 (early) detectable response defined as follows: (1) SI more than 10; or (2) ratio more than 2.5 of early harvest (D 2 or D 3) SI compared with pretreatment assay; or (3) ratio of more than 1 of early harvest to conventional (D 5) harvest.
IV indicates intravenous; SC, subcutaneous; inj, injection; and —, undetectable.
CFSE-based MLR assay with early harvest on D 3 and conventional D 5 harvest.
Thymidine-based MLR assay with early harvest on D 2 and conventional D 5 harvest.
Technical problem with incomplete CFSE labeling.
Technical difficulty: no third-party response or high self-background.