Table 6.
Cytokine release from human PBMC stimulated in vitro for 72 h in a solid phase assay with 1 μg well−1 of indicated therapeutic mAbs. Th1/Th2 7 and 10-Plex MSD® kits was used to measure IFNγ, TNFα, IL-1β, IL-2, IL-4, IL-5, IL-8, IL-10, IL-12 and IL-13 release. Cytokine ELISA was used to measure IL-22 and TGFβ release. The lectin PHA 10 μgml−1 was used as mitogen control. All responses were compared with IgG1 isotype control with the exception of TGN1412 which was compared with IgG4κ isotype control. Group responses shown are geometric mean followed by 95% confidence interval in brackets. Significant results are shown in bold (repeated measures analysis of log10 transformed data using Dunnett's method and 95% confidence). Donor responses shown (n = 7–16) are from 2–4 independent experiments
| Therapeutic mAb | IFNγ | TNFα | IL-1β | IL-2 | IL-4 | IL-5 |
|---|---|---|---|---|---|---|
| IgG1 control | 59.4 pg ml−1 (35.2, 100) | 726 pg ml−1 (494, 1066) | 30.9 pg ml−1 (20.0, 47.9) | 41.8 pg ml−1 (34.1, 51.1) | 9.8 pg ml−1 (8.8, 10.9) | 11.4 pg ml−1 (10.5, 12.3) |
| IgG4 control | 3.1 pg ml−1 (2.6, 3.6) | 25.4 pg ml−1 (16.3, 39.7) | 1.9 pg ml−1 (1.2, 3.2) | 9.5 pg ml−1 (7.2, 12.5) | 3.4 pg ml−1 (2.8, 4.0) | 3.7 pg ml−1 (2.2, 6.2) |
| Rituximab | 75.3 pg ml−1 (26.7, 212) | n.d. | n.d. | 33.3 pg ml−1 (26.5, 41.9) | 9.7 pg ml−1 (7.9, 11.8) | 11.5 pg ml−1 (9.7, 13.6) |
| Alemtuzumab | 88.8 pg ml−1 (43.3, 182) | 1972 pg ml−1 (1097, 3547) | 107 pg ml−1 (41.0, 280) | 41.7 pg ml−1 (35.2, 49.5) | 13.8 pg ml−1 (10.8, 17.7) | 14.6 pg ml−1 (11.6, 18.3) |
| Muromonab-CD3 | 18 013 pg ml−1 (13 598, 23 861) | 9855 pg ml−1 (7939, 12235) | 392 pg ml−1 (166, 926) | 2781 pg ml−1 (1453, 5325) | 41.0 pg ml−1 (31.1, 54.2) | 498 pg ml−1 (146, 1693) |
| TGN1412 | 30 748 pg ml−1 (20 767, 45 527) | 11314 pg ml−1 (10123, 12644) | 148 pg ml−1 (103, 212) | 8600 pg ml−1 (5229, 14144) | 54.0 pg ml−1 (46.6, 62.6) | 1904 pg ml−1 (1450, 2502) |
| Mitogen | 19 491 pg ml−1 (16 164, 23 503) | 6322 pg ml−1 (4953, 8070) | 289 pg ml−1 (194, 431) | 113 pg ml−1 (51.2, 247) | 35.9 pg ml−1 (29.4, 43.9) | 225 pg ml−1 (109, 465) |
| IL-8 | IL-10 | IL-12 | IL-13 | IL-22 | TGFβ | |
|---|---|---|---|---|---|---|
| IgG1 Control | 14 916 pg ml−1 (14 394, 15 457) | 28.8 pg ml−1 (25.8, 32.1) | 12.3 pg ml−1 (11.3, 13.4) | 16.0 pg ml−1 (12.9, 20.0) | 30.6 pg ml−1 (4.3, 219) | 1177 pg ml−1 (899, 1541) |
| IgG4 Control | 1 413 pg ml−1 (825, 2 423) | 11.1 pg ml−1 (8.3, 14.9) | 3.9 pg ml−1 (3.2, 4.7) | 2.4 pg ml−1 (1.8, 3.3) | 19.8 pg ml−1 (2.9, 132) | 715 pg ml−1 (560, 911) |
| Rituximab | n.d. | 26.7 pg ml−1 (21.0, 33.9) | 13.0 pg ml−1 (10.8, 15.6) | 17.8 pg ml−1 (12.5, 25.4) | 46.7 pg ml−1 (8.6, 254) | 1139 pg ml−1 (936, 1386) |
| Alemtuzumab | 15 218 pg ml−1 (14 482, 15 991) | 38.8 pg ml−1 (31.3, 48.0) | 17.5 pg ml−1 (13.3, 23.0) | 22.4 pg ml−1 (16.3, 30.9) | 37.0 pg ml−1 (6.5, 211) | 856 pg ml−1 (702, 1044) |
| Muromonab-CD3 | 15 445 pg ml−1 (14 604, 16 336) | 1060 pg ml−1 (695, 1616) | 35.1 pg ml−1 (25.6, 48.1) | 562 pg ml−1 (221, 1432) | 2870 pg ml−1 (1585, 5197) | 1036 pg ml−1 (840, 1277) |
| TGN1412 | 16 158 pg ml−1 (15 237, 17 134) | 240 pg ml−1 (166, 348) | 47.9 pg ml−1 (36.6, 62.7) | 1235 pg ml−1 (930, 1642) | 4743 pg ml−1 (2747, 8188) | 1050 pg ml−1 (743, 1484) |
| Mitogen | 15 132 pg ml−1 (14 741, 15 534) | 331 pg ml−1 (252, 435) | 44.3 pg ml−1 (30.6, 64.2) | 467 pg ml−1 (319, 683) | 872 pg ml−1 (522, 1457) | 1193 pg ml−1 (803, 1774) |