A rapid and specific whole blood HPA‐1 phenotyping by flow cytometry using two commercialized monoclonal antibodies directed against GP IIIa and GP IIb‐IIIa complexes

Sorel, Nathalie; Christiaens, Luc; Brizard, A.; Mauco, Gérard; Macchi, Laurent

doi:10.1046/j.1365-2141.2003.04757.x

Cited by 6 publications

(6 citation statements)

References 11 publications

(11 reference statements)

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“…1 Previous studies had demonstrated that MoAb SZ21 (anti-GPIIIa) bound markedly less to HPA-1b in comparison to HPA-1a PLTs 21 and could, therefore, be used for rapid PLT phenotyping of the HPA-1 system. 22 When we injected MoAb SZ21 over HPA-1a or HPA-1b sensor chips, similar interaction responses (approx. 100 RU) were detected during the association phase ( Fig.…”

Section: Spr Analysis Of Hpa-1a and -1b Antibodiesmentioning

confidence: 91%

Low‐avidity HPA‐1a alloantibodies in severe neonatal alloimmune thrombocytopenia are detectable with surface plasmon resonance technology

et al. 2009

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Section: Spr Analysis Of Hpa-1a and -1b Antibodiesmentioning

confidence: 91%

Low‐avidity HPA‐1a alloantibodies in severe neonatal alloimmune thrombocytopenia are detectable with surface plasmon resonance technology

et al. 2009

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“…SZ21, a monoclonal antibody (MoAb) directed against PLT glycoprotein (GP) IIIa, binds at or near the HPA-1a epitope 7-9 . Recently, we developed an in vivo model system in which human PLTs are injected into nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice and allowed to circulate for up to 24 hours 10 .…”

mentioning

confidence: 99%

“…SZ21, a monoclonal antibody (MoAb) directed against PLT glycoprotein (GP) IIIa, binds at or near the HPA-1a epitope. [7][8][9] Recently, we developed an in vivo model system in which human PLTs are injected into nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice and allowed to circulate for up to 24 hours. 10 Using this model, it is possible to investigate the fate of human PLTs in the presence of PLT-reactive antibodies and to examine the potential for novel therapeutics to prevent PLT clearance.…”

mentioning

confidence: 99%

Blockade of maternal anti‐HPA‐1a–mediated platelet clearance by an HPA‐1a epitope–specific F(ab′)₂ in an in vivo mouse model of alloimmune thrombocytopenia

et al. 2009

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Background Neonatal alloimmune thrombocytopenia (NAIT) is most commonly caused by transplacental passage of maternal human platelet-specific alloantigen (HPA)-1a antibodies that bind to fetal platelets (PLTs) and mediate their clearance. SZ21, a monoclonal antibody (MoAb) directed against PLT glycoprotein IIIa, competitively inhibits the binding of anti-HPA-1a alloantibodies to PLTs in vitro. The purpose of this investigation was to determine whether SZ21 F(ab′)2 fragments might be therapeutically effective in inhibiting or displacing maternal HPA-1a antibodies from the fetal PLT surface and preventing their clearance from circulation. Study Design and Methods Resting human PLTs from HPA-1ab heterozygous donors were injected into nonobese diabetic/severe combined immunodefi-cient (NOD/SCID) mice. Purified F(ab′)2 fragments of SZ21 or control immunoglobulin G (IgG) were injected intraperitoneally 30 minutes before introduction of HPA-1a antibodies. Blood samples were taken periodically and analyzed by flow cytometry to determine the percentage of circulating human PLTs. Results Anti-HPA-1a IgG from NAIT cases were able to efficiently clear HPA-1a–positive PLTs from murine circulation. Administration of SZ21 F(ab′)2 fragments not only inhibited binding of HPA-1a antibodies to circulating human PLTs, preventing their clearance, but also displaced bound HPA-1a antibodies from the PLT surface. Conclusion F(ab′)2 fragments of HPA-1a–selective MoAb SZ21 effectively inhibit anti-HPA-1a–mediated clearance of human PLT circulating in an in vivo NOD/SCID mouse model. These results suggest that agents that inhibit binding of anti-HPA-1a to PLTs may have therapeutic potential in the treatment of NAIT.

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“…5 Phenotyping of HPA 1 on platelets by flow cytometry has been reported. 12,13 Several molecular methods are also available for HPA genotyping.…”

mentioning

confidence: 99%

Genotyping of Human Platelet Antigens 1 to 6 and 15 by High-Resolution Amplicon Melting and Conventional Hybridization Probes

Liew

Nelson

Margraf

et al. 2006

The Journal of Molecular Diagnostics

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High-resolution melting techniques are a simple and cost-effective alternative to other closed-tube genotyping methods. Here, we genotyped human platelet antigens (HPAs) 1 to 6 and 15 by high-resolution melting methods that did not require labeled probes. Conventional melting analysis with hybridization probes (HybProbes) was also performed at each locus. HybProbe assays were performed individually, whereas amplicon melting (HPAs 1 to 5 and 16) and unlabeled probe (HPA 6) assays were duplexed when possible. At all loci for each method, both homozygous and heterozygous genotypes were easily identified. We analyzed 100 blinded clinical samples (33 amniotic fluid, 12 cultured amniocytes, and 55 blood samples) for all 7 single-nucleotide polymorphisms (SNPs) by each method. Genotype assignments could be made in 99.0% of the SNPs by high-resolution melting and in 98.7% of the SNPs with HybProbes with an overall genotype concordance of 98.8%. Errors included two sample misidentifications and six incorrect assignments that were all resolved by repeating the analysis. Advantages of high-resolution melting include rapid assay development and execution, no need for modified oligonucleotides, and similar accuracy in genotyping compared with other closed-tube melting methods. (J Mol

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A rapid and specific whole blood HPA‐1 phenotyping by flow cytometry using two commercialized monoclonal antibodies directed against GP IIIa and GP IIb‐IIIa complexes

Cited by 6 publications

References 11 publications

Low‐avidity HPA‐1a alloantibodies in severe neonatal alloimmune thrombocytopenia are detectable with surface plasmon resonance technology

Low‐avidity HPA‐1a alloantibodies in severe neonatal alloimmune thrombocytopenia are detectable with surface plasmon resonance technology

Blockade of maternal anti‐HPA‐1a–mediated platelet clearance by an HPA‐1a epitope–specific F(ab′)₂ in an in vivo mouse model of alloimmune thrombocytopenia

Genotyping of Human Platelet Antigens 1 to 6 and 15 by High-Resolution Amplicon Melting and Conventional Hybridization Probes

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A rapid and specific whole blood HPA‐1 phenotyping by flow cytometry using two commercialized monoclonal antibodies directed against GP IIIa and GP IIb‐IIIa complexes

Cited by 6 publications

References 11 publications

Low‐avidity HPA‐1a alloantibodies in severe neonatal alloimmune thrombocytopenia are detectable with surface plasmon resonance technology

Low‐avidity HPA‐1a alloantibodies in severe neonatal alloimmune thrombocytopenia are detectable with surface plasmon resonance technology

Blockade of maternal anti‐HPA‐1a–mediated platelet clearance by an HPA‐1a epitope–specific F(ab′)2 in an in vivo mouse model of alloimmune thrombocytopenia

Genotyping of Human Platelet Antigens 1 to 6 and 15 by High-Resolution Amplicon Melting and Conventional Hybridization Probes

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Blockade of maternal anti‐HPA‐1a–mediated platelet clearance by an HPA‐1a epitope–specific F(ab′)₂ in an in vivo mouse model of alloimmune thrombocytopenia