Effect of rhC3a and rhC5a on PAI-1 production in primary human skin mast cells, primary human blood basophils, HMC-1, and KU-812
| Cells . | Control . | rhC3a . | rhC5a . | Plasma C5a . |
|---|---|---|---|---|
| Primary human skin mast cells (healthy donors) | ||||
| Donor 1 | < 0.2 | ND | 5.3 ± 0.5 | ND |
| Donor 2 | < 0.2 | < 0.2 | 10.5 ± 1.6 | ND |
| Donor 3 | < 0.2 | < 0.2 | 7.9 ± 0.4 | 6.2 ± 0.7 |
| Donor 4 | < 0.2 | < 0.2 | 5.9 ± 0.4 | 4.6 ± 0.2 |
| Primary human blood basophils (healthy donors) | ||||
| Donor 1 | < 0.2 | < 0.2 | 1.4 ± 0.2 | ND |
| Donor 2 | < 0.2 | < 0.2 | 1.8 ± 0.2 | 1.6 ± 0.1 |
| Donor 3 | < 0.2 | < 0.2 | 1.2 ± 0.1 | 1.4 ± 0.1 |
| Primary human blood basophils (CML) | ||||
| Donor 1 | ||||
| Day 0 | < 0.2 | ND | 1.3 ± 0.1 | ND |
| Day 3 | < 0.2 | ND | 1.3 ± 0.2 | ND |
| Day 5 | < 0.2 | ND | 1.8 ± 0.1 | ND |
| Day 7 | < 0.2 | ND | 2.3 ± 0.3 | ND |
| Donor 2 | ||||
| Day 0 | < 0.2 | < 0.2 | 1.5 ± 0.2 | ND |
| Day 3 | < 0.2 | < 0.2 | 1.6 ± 0.1 | ND |
| Day 5 | < 0.2 | 0.3 ± 0.1 | 1.9 ± 0.1 | ND |
| Day 7 | < 0.2 | 0.3 ± 0.1 | 1.8 ± 0.3 | ND |
| Donor 3 | ||||
| Day 0 | < 0.2 | < 0.2 | 1.7 ± 0.3 | ND |
| Day 3 | < 0.2 | < 0.2 | 2.6 ± 0.2 | ND |
| Day 5 | ≪ 0.2 | 0.4 ± 0.1 | 2.1 ± 0.4 | ND |
| Day 7 | < 0.2 | 0.4 ± 0.2 | 3.5 ± 0.4 | ND |
| HMC-1 | < 0.2 | 0.6 ± 0.1 | 18.1 ± 1.9 | 16.3 ± 1.2 |
| KU-812 | 0.3 ± 0.1 | 0.4 ± 0.1 | 2.8 ± 0.4 | 4.1 ± 0.3 |
| Cells . | Control . | rhC3a . | rhC5a . | Plasma C5a . |
|---|---|---|---|---|
| Primary human skin mast cells (healthy donors) | ||||
| Donor 1 | < 0.2 | ND | 5.3 ± 0.5 | ND |
| Donor 2 | < 0.2 | < 0.2 | 10.5 ± 1.6 | ND |
| Donor 3 | < 0.2 | < 0.2 | 7.9 ± 0.4 | 6.2 ± 0.7 |
| Donor 4 | < 0.2 | < 0.2 | 5.9 ± 0.4 | 4.6 ± 0.2 |
| Primary human blood basophils (healthy donors) | ||||
| Donor 1 | < 0.2 | < 0.2 | 1.4 ± 0.2 | ND |
| Donor 2 | < 0.2 | < 0.2 | 1.8 ± 0.2 | 1.6 ± 0.1 |
| Donor 3 | < 0.2 | < 0.2 | 1.2 ± 0.1 | 1.4 ± 0.1 |
| Primary human blood basophils (CML) | ||||
| Donor 1 | ||||
| Day 0 | < 0.2 | ND | 1.3 ± 0.1 | ND |
| Day 3 | < 0.2 | ND | 1.3 ± 0.2 | ND |
| Day 5 | < 0.2 | ND | 1.8 ± 0.1 | ND |
| Day 7 | < 0.2 | ND | 2.3 ± 0.3 | ND |
| Donor 2 | ||||
| Day 0 | < 0.2 | < 0.2 | 1.5 ± 0.2 | ND |
| Day 3 | < 0.2 | < 0.2 | 1.6 ± 0.1 | ND |
| Day 5 | < 0.2 | 0.3 ± 0.1 | 1.9 ± 0.1 | ND |
| Day 7 | < 0.2 | 0.3 ± 0.1 | 1.8 ± 0.3 | ND |
| Donor 3 | ||||
| Day 0 | < 0.2 | < 0.2 | 1.7 ± 0.3 | ND |
| Day 3 | < 0.2 | < 0.2 | 2.6 ± 0.2 | ND |
| Day 5 | ≪ 0.2 | 0.4 ± 0.1 | 2.1 ± 0.4 | ND |
| Day 7 | < 0.2 | 0.4 ± 0.2 | 3.5 ± 0.4 | ND |
| HMC-1 | < 0.2 | 0.6 ± 0.1 | 18.1 ± 1.9 | 16.3 ± 1.2 |
| KU-812 | 0.3 ± 0.1 | 0.4 ± 0.1 | 2.8 ± 0.4 | 4.1 ± 0.3 |
Primary human skin mast cells from 4 different donors and primary human blood basophils from 3 different donors, isolated as described in “Materials and methods,” HMC-1 and KU-812 were incubated for 24 hours in the absence of presence of rhC3a or rhC5a or C5a purified from human plasma at a concentration of 1.0 μM, respectively. Human peripheral blood basophils were also isolated from 3 patients with CML and cultured for 7 days as described in “Materials and methods.” At days 0, 3, 5, and 7, aliquots of these cells were incubated for 24 hours in the absence or presence of the rhC5a at a concentration of 1.0 μM. Conditioned media of such treated cells were collected, and total PAI-1 antigen was determined as described in “Materials and methods.” Values are given in ng/106 cells per 24 hours and represent mean values ± SD of 3 independent determinations.
ND indicates not determined.