title: Anti-HPA-1a-mediated platelet phagocytosis by monocytes in vitro and its inhibition by Fc gamma receptor (FcgammaR) reactive reagents.
abstract: The study was undertaken to delineate mechanisms of platelet destruction by phagocytosis during fetal/neonatal alloimmune thrombocytopenia (FAIT/NAIT) because of maternal antibodies against human platelet antigen 1a (HPA-1a). By employing a platelet phagocytosis assay based on the ORPEGEN flow cytometric bacterial phagocytosis test, we measured monocyte ingestion of platelets mediated by anti-HPA-1a antibodies. Moreover, we tested, as potential therapeutic agents, FcgammaR reactive reagents, for their inhibition of this process. Four of six anti-HPA-1a sera tested mediated phagocytosis of HPA-1a-positive platelets in a concentration-dependent manner. Monocyte ingestion of platelets was almost completely inhibited by cytochalasin D. No anti-HPA-1a-mediated phagocytosis was observed with anti-HPA-1a-negative platelets. The humanised anti-FcgammaRI monoclonal antibody H22 at concentrations 1-100 microg/ml, completely inhibited anti-HPA-1a-mediated phagocytosis as did similar concentrations of ivIg. By contrast, a mouse monoclonal anti-FcgammaRII (IV.3, Fab) at 10 microg/ml caused little or no suppression of platelet phagocytosis mediated by two anti-HPA-1 sera. Furthermore, the addition of anti-FcgammaRII (10 microg/ml) to sub-optimal concentrations of H22 did not significantly increase the inhibitory effect of the latter compound. Monomeric IgG (0.1-10 microg/ml) failed to suppress anti-HPA-1 mediated platelet ingestion by the phagocytes, as did anti-FcgammaRIII. To our knowledge this is a rare example of an assay that measures platelet phagocytosis in vitro. The results suggest that FcgammaRI plays a major role in anti-HPA-1a-mediated platelet phagocytosis by monocytes while FcgammaRIIa, is of little or minor importance only. Moreover, the findings indicate the use of H22 as an alternative to interavenous Ig (ivIg) in the management of FAIT/NAIT.
authors.affiliation: Dept of Haematology, St Mary's, Imperial College Faculty of Medicine, London. e.wiener@ic.ac.uk
authors.forename: E
authors.initials: E
authors.lastname: Wiener
authors.forename: O
authors.initials: O
authors.lastname: Abeyakoon
authors.forename: G
authors.initials: G
authors.lastname: Benchetrit
authors.forename: M
authors.initials: M
authors.lastname: Lyall
authors.forename: T
authors.initials: T
authors.lastname: Keler
authors.forename: C H
authors.initials: CH
authors.lastname: Rodeck
mesh_terms: D000328:Adult; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D016824:Antigens, Human Platelet / Q000276:immunology*; D001792:Blood Platelets / Q000276:immunology*; D003601:Cytotoxicity Tests, Immunologic; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007202:Indicators and Reagents / Q000494:pharmacology; D007231:Infant, Newborn; D007232:Infant, Newborn, Diseases / Q000276:immunology; D039661:Integrin beta3; D008431:Maternal-Fetal Exchange / Q000276:immunology; D009000:Monocytes / Q000276:immunology*; D010587:Phagocytosis / Q000187:drug effects*; D011247:Pregnancy; D017452:Receptors, IgG / Q000276:immunology*; D013921:Thrombocytopenia / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D016824:Antigens, Human Platelet; C061248:ITGB3 protein, human; D007202:Indicators and Reagents; D039661:Integrin beta3; D017452:Receptors, IgG