Table 1.
Comparative analysis of IL-8, IL-13 and IL-17 release with different CRA methods: Cytokine release expressed in pg ml− 1 from human PBMC SP (1 μg well− 1 coating concentration), PBMC HDC (1 μg ml− 1), WB and 10% (v/v) WB CRA (5 μg ml− 1) stimulated with the indicated therapeutic mAbs after 48 h stimulation, with the exception of the PBMC HDC CRA which utilizes a 48 h HDC followed by 24 h stimulation with mAbs. PHA at 10 μg ml− 1 was used as a mitogen control. All responses were compared against IgG1 isotype control with the exception of TGN1412, which was compared against IgG4κ isotype control. Group responses shown are the geometric means followed by the 95% confidence intervals in brackets. Significant results (p < 0.05) are shown in bold (repeated measures analysis of log10 transformed data using Tukey's method with 95.0% confidence). Donor responses shown are n = 10 from 3 independent experiments, except for the PBMC HDC CRA for which n = 8 from 2 independent experiments. CRA comparisons were performed using the same set of donors, except for the PBMC HDC CRA which utilized a different set of 8 donors.
| Cytokine | CRA method | IgG1 control | IgG4 control | Rituximab | Alemtuzumab | Muromonab-CD3 | TGN1412 | CD28 agonist | Mitogen control |
|---|---|---|---|---|---|---|---|---|---|
| IL-8 | PBMC SP | 9202 (8696, 9738) | 7066 (6206, 8045) | 7520 (6803, 8313) | 7648 (7088, 8254) | 193,381 (150,646, 248,239) | 136,048 (91,305, 202,716) | 9291 (2934, 29,415) | 210,755 (156,467, 283,879) |
| PBMC HDC | 3566 (2715, 4683) | 3156 (2098, 4746) | 1117 (53, 23383) | 1137 (54, 23921) | 1148(54, 24,265) | 1137 (54, 23,946) | 2510 (1298, 4854) | 4112 (3279, 5155) | |
| WB | 2102 (879, 5028) | 2538 (1038, 6205) | 1815 (896, 3676) | 7162 (5033, 10192) | 1256 (814, 1938) | 2474 (842, 7274) | 1504 (882, 2566) | 19,529 (6838, 55,771) | |
| 10% (v/v) WB | 188 (88, 400) | 252 (79, 806) | 82 (41, 163) | 302 (224, 408) | 2855 (1156, 7048) | 83 (41, 170) | 130 (55, 304) | 41,750 (20,714, 84,151) | |
| IL-13 | PBMC SP | 140 (91, 213) | 67 (45, 101) | 136 (112, 166) | 192 (113, 326) | 1625 (1289, 2049) | 2451 (1719, 3494) | 300 (172, 521) | 1932 (1402, 2662) |
| PBMC HDC | 119 (80, 176) | 101 (66, 155) | 94 (24, 364) | 96 (24, 383) | 184 (38, 899) | 293 (55, 1567) | 67 (31, 147) | 779 (553, 1098) | |
| WB | 13 (6.6, 24) | 15 (7.6, 28) | 16 (11, 24) | 39 (27, 56) | 9.7 (3.7, 25) | 19 (5.4, 68) | 31 (14, 68) | 224 (102, 493) | |
| 10% (v/v) WB | 2.9 (1.8, 4.9) | 3.6 (1.8, 7.3) | 2 (1.5, 2.5) | 3 (2.3, 3.9) | 26 (6.8, 100) | 6.4 (2.3, 18) | 41 (22, 77) | 372 (218, 635) | |
| IL-17 | PBMC SP | 1.3 (0.75, 2.1) | 2.3 (0.66, 7.7) | 1.6 (0.79, 3.1) | 1.9 (0.72, 5) | 629 (365, 1085) | 346 (123, 974) | 1.7 (0.75, 3.9) | 71 (36, 138) |
| PBMC HDC | 1.5 (0.58, 3.8) | 2.7 (0.83, 9) | 3.5 (0.8, 16) | 3.5 (0.77, 16) | 1.6 (0.55, 4.4) | 3 (0.54, 17) | 2.7 (0.84, 8.5) | 6.4 (1.1, 37) | |
| WB | 1.6 (0.78, 3.3) | 1.5 (0.8, 2.8) | 1.4 (0.64, 3.2) | 1.4 (0.65, 3) | 1.4 (0.63, 3.3) | 1 (1, 1) | 2 (0.87, 4.7) | 1.2 (0.77, 2) | |
| 10% (v/v) WB | 1.3 (0.71, 2.4) | 4.1 (1.3, 13) | 1.8 (0.73, 4.4) | 1.7 (0.73, 4.2) | 3.5 (1.1, 11) | 2.2 (0.86, 5.9) | 2.5 (0.84, 7.3) | 152 (42, 552) |