Platelet activation in newborns detected by flow-cytometry

Platelet function was investigated in full-term infants on the first, the fourth and tenth days of life and compared to normal adult controls. Platelet function was analyzed through a new cytofluorimetric technique with two murine monoclonal antibodies, PAC-1 and anti-GMP-140, directed against two membrane proteins expressed on the activated platelets' surface. The percentage of activated platelets detected with PAC-1 and anti-GMP-140 was evaluated at basal condition and after in vitro stimulation with a weak agonist (ADP) and a strong Txa2 analogue inducer (U 46619). At day 1 platelet activation at basal condition was negligible and similar to adult controls both with PAC-1 (1.2 vs 1.1%) and anti-GMP-140 (2.6 vs. 3.3%). On the contrary, after ADP stimulation the percentage of PAC-1-positive activated platelets was significantly reduced in neonates compared to adults (22 vs. 66%; p < 0.001) and even more after U 46619 (11 vs. 72%; p < 0.001). The percentage of anti-GMP-140-positive activated platelets behaved similarly after adding both ADP (26 vs. 46%; p < 0.01) and U 46619 (37 vs. 67%; p < 0.001). The reduced platelet activation after ADP and U 46619 persisted at day 4 both with PAC-1 and with anti-GMP-140. On the contrary, at day 10 newborn platelets analyzed with anti-GMP-140 behaved similarly to the adult ones both at basal condition and after stimulation with ADP or U 46619 (6 vs. 3% at basal state, 42 vs. 46% after ADP addition, and 55 vs. 67% after U 46619). These data demonstrate that the reduced platelet activation present in newborns is restored by the tenth day after birth.