Effects of mAbs against PSGL-1, P-selectin, or platelets, rmP-selectin–Fc fusion protein, and heparin on leukocyte rolling, leukocyte-platelet adhesion, and blood neutrophil counts before and after IM7 injection
| Treatment . | Rolling cells, % of baseline . | Cells with platelets, % of total . | Neutrophil counts, % of baseline . | |||
|---|---|---|---|---|---|---|
| Median (range) . | Median (range) . | Median (range) . | ||||
| 25 min . | +IM7 (5 min) . | 25 min . | +IM7 (5 min) . | 25 min . | +IM7 (5 min) . | |
| PBS (n = 5 mice) | 114 (91-116) | 2 (0-5) | 2 (0-4) | 91 (86-100) | 94 (86-110) | 11 (4-23) |
| Rat IgG (n = 5 mice) | 96 (93-104) | 0 (0-2) | 1 (0-3) | 95 (91-99) | 106 (97-121) | 17 (7-23) |
| Anti-PSGL (n = 3 mice) | 47 (32-70) | 5 (2-10) | 1 (0-1) | 82 (70-86) | 113 (100-115) | 20 (11-34) |
| Anti–P-sel (n = 3 mice) | 88 (81-98) | 9 (0-15) | 11 (6-19) | 40 (21-58) | 85 (69-89) | 25 (6-31) |
| rmP-sel–Fc (n = 3 mice) | 3 (2-8) | 0 (0-1) | 98 (97-100) | 100 (98-100) | 13 (8-33) | 10 (7-19) |
| Antiplatelet (n = 5 mice) | 95 (90-106) | 16 (3-27) | 2 (1-6) | 1 (0-5) | 97 (75-128) | 31 (20-45) |
| Heparin (n = 5 mice) | 76 (54-84) | 24 (18-49) | 0 (0-1) | 4 (2-17) | 120 (83-126) | 37 (29-60) |
| Treatment . | Rolling cells, % of baseline . | Cells with platelets, % of total . | Neutrophil counts, % of baseline . | |||
|---|---|---|---|---|---|---|
| Median (range) . | Median (range) . | Median (range) . | ||||
| 25 min . | +IM7 (5 min) . | 25 min . | +IM7 (5 min) . | 25 min . | +IM7 (5 min) . | |
| PBS (n = 5 mice) | 114 (91-116) | 2 (0-5) | 2 (0-4) | 91 (86-100) | 94 (86-110) | 11 (4-23) |
| Rat IgG (n = 5 mice) | 96 (93-104) | 0 (0-2) | 1 (0-3) | 95 (91-99) | 106 (97-121) | 17 (7-23) |
| Anti-PSGL (n = 3 mice) | 47 (32-70) | 5 (2-10) | 1 (0-1) | 82 (70-86) | 113 (100-115) | 20 (11-34) |
| Anti–P-sel (n = 3 mice) | 88 (81-98) | 9 (0-15) | 11 (6-19) | 40 (21-58) | 85 (69-89) | 25 (6-31) |
| rmP-sel–Fc (n = 3 mice) | 3 (2-8) | 0 (0-1) | 98 (97-100) | 100 (98-100) | 13 (8-33) | 10 (7-19) |
| Antiplatelet (n = 5 mice) | 95 (90-106) | 16 (3-27) | 2 (1-6) | 1 (0-5) | 97 (75-128) | 31 (20-45) |
| Heparin (n = 5 mice) | 76 (54-84) | 24 (18-49) | 0 (0-1) | 4 (2-17) | 120 (83-126) | 37 (29-60) |
Mice were pretreated intravenously with PBS or one of the listed reagents 25 minutes before IM7 (200 μg intravenously) administration. Rat IgG, mAbs against PSGL-1, and P-selectin were injected at 200 μg/mouse. rmP-selectin–Fc was given at a dose of 30 μg, antiplatelet pAb at 100 μg, and heparin at 40 U/mouse (optimum doses were established in preliminary experiments). Mice were subjected to IVM before and 25 minutes after the pretreatment and 5 minutes after IM7 injection (+IM7), and were also bled for cell counts at the same time points (during the video recordings). The number of rolling cells is expressed as the percentage of baseline (at 0 minutes). Platelet binding is expressed as the percentage of adherent cells with surface-associated platelets, and neutrophil counts as the percentage of baseline values.