Neutrophils depend upon phosphatidylserine exposure to phagocytose activated platelets
. | Neutrophils with adherent platelets, % . | Neutrophils with internalized platelets, % . |
---|---|---|
Resting platelets | 1.7 ± 0.8 | 0.3 ± 0.2 |
Activated platelets | 25.6 ± 3.0 | 37.1 ± 4.4 |
Activated platelets + trypsin-EDTA | 9.7 ± 0.8* | 0.1 ± 15.2 |
Annexin A5–treated activated platelets | 20.1 ± 3.3 | 5.1 ± 1.2* |
Annexin A5–treated activated platelets + trypsin-EDTA | 7.1 ± 3.7* | 2.0 ± 1.2* |
. | Neutrophils with adherent platelets, % . | Neutrophils with internalized platelets, % . |
---|---|---|
Resting platelets | 1.7 ± 0.8 | 0.3 ± 0.2 |
Activated platelets | 25.6 ± 3.0 | 37.1 ± 4.4 |
Activated platelets + trypsin-EDTA | 9.7 ± 0.8* | 0.1 ± 15.2 |
Annexin A5–treated activated platelets | 20.1 ± 3.3 | 5.1 ± 1.2* |
Annexin A5–treated activated platelets + trypsin-EDTA | 7.1 ± 3.7* | 2.0 ± 1.2* |
Neutrophils effectively adhere to and phagocytose activated but not resting platelets after a 5-minute interaction at 37°C. Treatment with trypsin-EDTA, which acts in the extracellular environment, strips adherent but not internalized platelets (*P < .05 by paired t test). Hindrance with phosphatidylserine recognition by annexin A5 blocks platelet internalization but does not influence adhesion (*P < .05 by paired t test). Both platelet adhesion and internalization were nondetectable when (1) phagocytosis was blocked by annexin A5 and (2) extracellular platelets were removed after the assay by trypsin-EDTA (*P < .05 by paired t test). Platelet adherence and internalization were determined by flow cytometry, as described in “Adhesion and phagocytosis.” Results are expressed as mean ± SEM of 3 independent experiments performed in duplicate.