Current Anti-HPA-1a Standard Antibodies React with the β3 Integrin Subunit but not with αIIbβ3 and αvβ3 Complexes
| Autor: | Sentot Santoso, Behnaz Bayat, Silke Werth, Annalena Traum, Jieging Zhu, Heike Berghöfer, Gregor Bein, Ulrich J. Sachs |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
medicine.drug_class Neovascularization Physiologic Platelet Glycoprotein GPIIb-IIIa Complex Immunologic Tests 030204 cardiovascular system & hematology Monoclonal antibody Severity of Illness Index Article 03 medical and health sciences 0302 clinical medicine Antigen Antibody Specificity Isoantibodies Predictive Value of Tests medicine Humans Antigens Human Platelet Platelet Tube formation biology Chemistry Integrin beta3 Antibodies Monoclonal Endothelial Cells Reproducibility of Results Hematology Transfection Integrin alphaVbeta3 medicine.disease Molecular biology Thrombocytopenia Neonatal Alloimmune HEK293 Cells 030104 developmental biology Cell culture Neonatal alloimmune thrombocytopenia biology.protein Antibody |
| Zdroj: | Thromb Haemost |
| ISSN: | 2567-689X 0340-6245 |
| DOI: | 10.1055/s-0039-1696716 |
| Popis: | Background Fetal/neonatal alloimmune thrombocytopenia (FNAIT) results from maternal alloantibodies (abs) reacting with fetal platelets expressing paternal human platelet antigens (HPAs), mostly HPA-1a. Anti-HPA-1a abs, are the most frequent cause of severe thrombocytopenia and intracranial hemorrhage (ICH). Objectives Titration of anti-HPA-1a in maternal serum using standard National Institute for Biological Standards and Control (NIBSC) 03/152 is one diagnostic approach to predict the severity of FNAIT. Recently, we found three anti-HPA-1a subtypes reacting with the β3 subunit independently or dependently from complexes with αIIb and αv. Endothelial cell-reactive anti-αvβ3 abs were found predominantly in cases with ICH. Our aim was to assess whether available standard material represents all anti-HPA-1a subtypes. Materials and Methods In this study, anti-HPA-1a sera (NIBSC 03/152) and human monoclonal antibodies (moabs) against HPA-1a (moabs 26.4 and 813) were evaluated using transfected cell lines expressing αIIbβ3, αvβ3 or monomeric cβ3. Results Flow cytometry analyses with well-characterized murine moabs recognizing αIIbβ3, αvβ3, or β3 alone demonstrated that AP3 reacts compound-independently, whereas compound-dependent moabs Gi5 and 23C6 reacted only with complexes. NIBSC 03/152, moabs 26.4, and 813 against HPA-1a reacted like AP3, same results were obtained with monomeric cβ3 in immunoblotting. Antigen capture assay targeting endothelial cells showed anti-HPA-1a reactivity disappearance after cβ3 beads adsorption. Furthermore, in contrast to anti-HPA-1a abs from ICH cases, none of NIBSC 03/152, 26.4, and 813 inhibited tube formation. Conclusion These results suggest that current anti-HPA-1a standard material contains only the anti-β3 subtype. The absence of anti-αvβ3 makes NIBSC 03/152 less suitable as standard to predict the severity of FNAIT. |
| Databáze: | OpenAIRE |
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