Assessment of anti-CD52 on the development of moDCs in vitro
| . | Cell no. . | Apoptotic cells . | CD83+ cells d8, % . | Stimulatory capacity alloMLR, % . | ||
|---|---|---|---|---|---|---|
| d6 relative to d 0, % . | d8 relative to d 0, % . | d 6, % . | d 8, % . | |||
| Anti-CD52 (alemtuzumab) | 104 | 105 | 5 | 10 | 94 | 99 |
| Anti-CD20 negative control (rituximab) | 100 | 100 | 8 | 20 | 92 | 100 |
| . | Cell no. . | Apoptotic cells . | CD83+ cells d8, % . | Stimulatory capacity alloMLR, % . | ||
|---|---|---|---|---|---|---|
| d6 relative to d 0, % . | d8 relative to d 0, % . | d 6, % . | d 8, % . | |||
| Anti-CD52 (alemtuzumab) | 104 | 105 | 5 | 10 | 94 | 99 |
| Anti-CD20 negative control (rituximab) | 100 | 100 | 8 | 20 | 92 | 100 |
In vitro cultures for the generation of moDC from CD14+ monocytes were supplemented with humanized anti-CD52 (alemtuzumab) or anti-CD20 (rituximab) as a negative control, both in excess at 1 mg/mL. Viable cell numbers were based on trypan blue exclusion by direct hemacytometer counts and were expressed as a percentage relative to the starting number of monocytes at day 0. Annexin V and propidium iodide staining to identify the percent apoptotic cells in culture did not reveal any substantial differences relative to the negative control MAb. Staining for CD83, as well as CD40, CD80, and CD86 (not shown) confirmed efficient maturation in both conditions. Proliferation of responder T cells in alloMLR showed equal stimulatory capacity for moDC generated in the presence of alemtuzumab compared with the negative control.