Antiendothelial αvβ3 Antibodies Are a Major Cause of Intracranial Bleeding in Fetal/Neonatal Alloimmune Thrombocytopenia.
Fetal/neonatal alloimmune thrombocytopenia is a severe bleeding disorder, which can result in intracranial hemorrhage (ICH), leading to death or neurological sequelae. In whites, maternal anti-human platelet antigen-1a (HPA-1a) antibodies are responsible for the majority of cases. No predictive factors for ICH are available to guide prophylactic treatment during pregnancy. In this study, we investigated antibodies from mothers with ICH-positive fetal/neonatal alloimmune thrombocytopenia and with ICH-negative fetal/neonatal alloimmune thrombocytopenia to identify serological and functional differences between the groups.



In an antigen capture assay, we observed a stronger binding of +ICH antibodies to endothelial cell (EC)-derived αvβ3. By absorption experiments, we subsequently identified anti-HPA-1a antibodies of anti-αvβ3 specificity in the +ICH but not in the -ICH cohort. Only the anti-αvβ3 subtype, but not the anti-β3 subtype, induced EC apoptosis of HPA-1a-positive ECs by caspase-3/7 activation, and mediated by reactive oxygen species. In addition, only the anti-αvβ3 subtype, but not the anti-β3 subtype, interfered with EC adhesion to vitronectin and with EC tube formation.



We conclude that the composition of the anti-HPA-1a antibody subtype(s) of the mother may determine whether ICH occurs. Analysis of anti-HPA-1a antibodies of the anti-αvβ3 subtype in maternal serum has potential in the diagnostic prediction of ICH development and may allow for modification of prophylactic treatment in fetal/neonatal alloimmune thrombocytopenia.
From the Institute for Clinical Immunology and Transfusion Medicine, Justus Liebig University, Giessen, Germany (S.S., H.W., S.W., G.B., B.B., U.J.S.); Department of Anatomy, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia (H.W.); Institute for Immunology and Transfusion Medicine, Ernst-Moritz-Arndt University, Greifswald, Germany (T.B.); Center for Transfusion Medicine and Hemotherapy (N.A.-F., U.J.S.) and German Center for Fetomaternal Incompatibility (DZFI) (G.B., U.J.S.), University Hospital Giessen and Marburg, Giessen, Germany; Institute for Transfusion Medicine, University of Rostock, Rostock, Germany (V.K.); and Blood Research Institute, BloodCenter of Wisconsin, Milwaukee (J.Z., P.J.N.). sentot.santoso@immunologie.med.uni-giessen.de ulrich.sachs@med.uni-giessen.de.
Sentot
S
Santoso
From the Institute for Clinical Immunology and Transfusion Medicine, Justus Liebig University, Giessen, Germany (S.S., H.W., S.W., G.B., B.B., U.J.S.); Department of Anatomy, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia (H.W.); Institute for Immunology and Transfusion Medicine, Ernst-Moritz-Arndt University, Greifswald, Germany (T.B.); Center for Transfusion Medicine and Hemotherapy (N.A.-F., U.J.S.) and German Center for Fetomaternal Incompatibility (DZFI) (G.B., U.J.S.), University Hospital Giessen and Marburg, Giessen, Germany; Institute for Transfusion Medicine, University of Rostock, Rostock, Germany (V.K.); and Blood Research Institute, BloodCenter of Wisconsin, Milwaukee (J.Z., P.J.N.).
Hevi
H
Wihadmadyatami
From the Institute for Clinical Immunology and Transfusion Medicine, Justus Liebig University, Giessen, Germany (S.S., H.W., S.W., G.B., B.B., U.J.S.); Department of Anatomy, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia (H.W.); Institute for Immunology and Transfusion Medicine, Ernst-Moritz-Arndt University, Greifswald, Germany (T.B.); Center for Transfusion Medicine and Hemotherapy (N.A.-F., U.J.S.) and German Center for Fetomaternal Incompatibility (DZFI) (G.B., U.J.S.), University Hospital Giessen and Marburg, Giessen, Germany; Institute for Transfusion Medicine, University of Rostock, Rostock, Germany (V.K.); and Blood Research Institute, BloodCenter of Wisconsin, Milwaukee (J.Z., P.J.N.).
Tamam
T
Bakchoul
From the Institute for Clinical Immunology and Transfusion Medicine, Justus Liebig University, Giessen, Germany (S.S., H.W., S.W., G.B., B.B., U.J.S.); Department of Anatomy, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia (H.W.); Institute for Immunology and Transfusion Medicine, Ernst-Moritz-Arndt University, Greifswald, Germany (T.B.); Center for Transfusion Medicine and Hemotherapy (N.A.-F., U.J.S.) and German Center for Fetomaternal Incompatibility (DZFI) (G.B., U.J.S.), University Hospital Giessen and Marburg, Giessen, Germany; Institute for Transfusion Medicine, University of Rostock, Rostock, Germany (V.K.); and Blood Research Institute, BloodCenter of Wisconsin, Milwaukee (J.Z., P.J.N.).
Silke
S
Werth
From the Institute for Clinical Immunology and Transfusion Medicine, Justus Liebig University, Giessen, Germany (S.S., H.W., S.W., G.B., B.B., U.J.S.); Department of Anatomy, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia (H.W.); Institute for Immunology and Transfusion Medicine, Ernst-Moritz-Arndt University, Greifswald, Germany (T.B.); Center for Transfusion Medicine and Hemotherapy (N.A.-F., U.J.S.) and German Center for Fetomaternal Incompatibility (DZFI) (G.B., U.J.S.), University Hospital Giessen and Marburg, Giessen, Germany; Institute for Transfusion Medicine, University of Rostock, Rostock, Germany (V.K.); and Blood Research Institute, BloodCenter of Wisconsin, Milwaukee (J.Z., P.J.N.).
Nadia
N
Al-Fakhri
From the Institute for Clinical Immunology and Transfusion Medicine, Justus Liebig University, Giessen, Germany (S.S., H.W., S.W., G.B., B.B., U.J.S.); Department of Anatomy, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia (H.W.); Institute for Immunology and Transfusion Medicine, Ernst-Moritz-Arndt University, Greifswald, Germany (T.B.); Center for Transfusion Medicine and Hemotherapy (N.A.-F., U.J.S.) and German Center for Fetomaternal Incompatibility (DZFI) (G.B., U.J.S.), University Hospital Giessen and Marburg, Giessen, Germany; Institute for Transfusion Medicine, University of Rostock, Rostock, Germany (V.K.); and Blood Research Institute, BloodCenter of Wisconsin, Milwaukee (J.Z., P.J.N.).
Gregor
G
Bein
From the Institute for Clinical Immunology and Transfusion Medicine, Justus Liebig University, Giessen, Germany (S.S., H.W., S.W., G.B., B.B., U.J.S.); Department of Anatomy, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia (H.W.); Institute for Immunology and Transfusion Medicine, Ernst-Moritz-Arndt University, Greifswald, Germany (T.B.); Center for Transfusion Medicine and Hemotherapy (N.A.-F., U.J.S.) and German Center for Fetomaternal Incompatibility (DZFI) (G.B., U.J.S.), University Hospital Giessen and Marburg, Giessen, Germany; Institute for Transfusion Medicine, University of Rostock, Rostock, Germany (V.K.); and Blood Research Institute, BloodCenter of Wisconsin, Milwaukee (J.Z., P.J.N.).
Volker
V
Kiefel
From the Institute for Clinical Immunology and Transfusion Medicine, Justus Liebig University, Giessen, Germany (S.S., H.W., S.W., G.B., B.B., U.J.S.); Department of Anatomy, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia (H.W.); Institute for Immunology and Transfusion Medicine, Ernst-Moritz-Arndt University, Greifswald, Germany (T.B.); Center for Transfusion Medicine and Hemotherapy (N.A.-F., U.J.S.) and German Center for Fetomaternal Incompatibility (DZFI) (G.B., U.J.S.), University Hospital Giessen and Marburg, Giessen, Germany; Institute for Transfusion Medicine, University of Rostock, Rostock, Germany (V.K.); and Blood Research Institute, BloodCenter of Wisconsin, Milwaukee (J.Z., P.J.N.).
Jieqing
J
Zhu
From the Institute for Clinical Immunology and Transfusion Medicine, Justus Liebig University, Giessen, Germany (S.S., H.W., S.W., G.B., B.B., U.J.S.); Department of Anatomy, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia (H.W.); Institute for Immunology and Transfusion Medicine, Ernst-Moritz-Arndt University, Greifswald, Germany (T.B.); Center for Transfusion Medicine and Hemotherapy (N.A.-F., U.J.S.) and German Center for Fetomaternal Incompatibility (DZFI) (G.B., U.J.S.), University Hospital Giessen and Marburg, Giessen, Germany; Institute for Transfusion Medicine, University of Rostock, Rostock, Germany (V.K.); and Blood Research Institute, BloodCenter of Wisconsin, Milwaukee (J.Z., P.J.N.).
Peter J
PJ
Newman
From the Institute for Clinical Immunology and Transfusion Medicine, Justus Liebig University, Giessen, Germany (S.S., H.W., S.W., G.B., B.B., U.J.S.); Department of Anatomy, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia (H.W.); Institute for Immunology and Transfusion Medicine, Ernst-Moritz-Arndt University, Greifswald, Germany (T.B.); Center for Transfusion Medicine and Hemotherapy (N.A.-F., U.J.S.) and German Center for Fetomaternal Incompatibility (DZFI) (G.B., U.J.S.), University Hospital Giessen and Marburg, Giessen, Germany; Institute for Transfusion Medicine, University of Rostock, Rostock, Germany (V.K.); and Blood Research Institute, BloodCenter of Wisconsin, Milwaukee (J.Z., P.J.N.).
Behnaz
B
Bayat
From the Institute for Clinical Immunology and Transfusion Medicine, Justus Liebig University, Giessen, Germany (S.S., H.W., S.W., G.B., B.B., U.J.S.); Department of Anatomy, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia (H.W.); Institute for Immunology and Transfusion Medicine, Ernst-Moritz-Arndt University, Greifswald, Germany (T.B.); Center for Transfusion Medicine and Hemotherapy (N.A.-F., U.J.S.) and German Center for Fetomaternal Incompatibility (DZFI) (G.B., U.J.S.), University Hospital Giessen and Marburg, Giessen, Germany; Institute for Transfusion Medicine, University of Rostock, Rostock, Germany (V.K.); and Blood Research Institute, BloodCenter of Wisconsin, Milwaukee (J.Z., P.J.N.). sentot.santoso@immunologie.med.uni-giessen.de ulrich.sachs@med.uni-giessen.de.
Ulrich J
UJ
Sachs
D000818:Animals; D000918:Antibody Specificity; D016824:Antigens, Human Platelet / Q000276:immunology / Q000378:metabolism; D017209:Apoptosis; D001323:Autoantibodies / Q000276:immunology* / Q000378:metabolism; D016466:CHO Cells; D016022:Case-Control Studies; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D002448:Cell Adhesion; D003412:Cricetulus; D042783:Endothelial Cells / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D005260:Female; D005865:Gestational Age; D061307:Human Umbilical Vein Endothelial Cells / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007231:Infant, Newborn; D039302:Integrin alphaVbeta3 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D039661:Integrin beta3; D020300:Intracranial Hemorrhages / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D008297:Male; D008431:Maternal-Fetal Exchange; D018919:Neovascularization, Physiologic; D011247:Pregnancy; D017382:Reactive Oxygen Species / Q000378:metabolism; D054098:Thrombocytopenia, Neonatal Alloimmune / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D014162:Transfection
D016824:Antigens, Human Platelet; D001323:Autoantibodies; C061248:ITGB3 protein, human; D039302:Integrin alphaVbeta3; D039661:Integrin beta3; D017382:Reactive Oxygen Species; C505407:CASP3 protein, human; C505520:CASP7 protein, human; D053148:Caspase 3; D053179:Caspase 7
antibodies; endothelial cells; intracranial hemorrhages; reactive oxygen species; thrombocytopenia, neonatal alloimmune