Functional heterogeneity of alloantibodies against the human platelet antigen (HPA)-1a

Abstract

The integrin alphaIIbbeta is the major fibrinogen receptor on the platelet membrane and plays a crucial role for platelet aggregation. The beta3-subunit carries the human platelet alloantigen (HPA)-1a, which is the main target for alloantibodies (alloabs) responsible for foetal and neonatal alloimmune thrombocytopenia (FNAIT) and post-transfusion purpura (PTP). Whereas PTP is almost invariably associated with severe bleeding, the clinical presentation of FNAIT ranges from mild thrombocytopenia to severe haemorrhagic diathesis. However, this clinical heterogeneity is not fully understood as it is not explained solely by the variability of the platelet count. Here, we examined the ability of HPA-1a alloabs from mothers with FNAIT (n = 43) and PTP patients (n = 8) to inhibit cell adhesion to fibrinogen and asked if this inhibition was correlated with the heterogeneity of the clinical picture. Stably transfected cells expressing HPA-1a (beta3-Leu33) and -1b (beta3-Pro33) isoforms were incubated with sera containing HPA-1a alloabs and were allowed to adhere to immobilised fibrinogen. The inhibitory activity was measured as percentage of cell adhesion in the presence of patient sera versus normal AB serum. Only two FNAIT sera specifically inhibited the adhesion of HPA-1a, but not HPA-1b cells. Two other FNAIT sera blocked the adhesion of HPA-1a as well as HPA-1b cells. Interestingly, all four neonates with inhibitory HPA-1a alloabs (9% of all sera) suffered severe bleeding. In comparison, the majority of PTP sera (75%) inhibited cell binding to fibrinogen, four PTP sera selectively inhibited the adhesion of HPA-1a cells whilst 2 sera impaired the binding of both allotypes. Our observations indicate that 1) HPA-1a alloabs are heterogeneous in their ability to interfere with fibrinogen binding, and 2) inhibition of the alphaIIbbeta3 fibrinogen receptor by HPA-1a alloabs may contribute to pronounced bleeding in patients with alloimmune thrombocytopenia.