Recombinant Anti-PF4 Antibodies Derived from Patients with Vaccine-Induced Immune Thrombocytopenia and Thrombosis (VITT) Facilitate Research and Laboratory Diagnosis of VITT
Background/Objectives: Adenoviral vector-based vaccines against COVID-19 rarely cause vaccine-induced immune thrombocytopenia and thrombosis (VITT), a severe adverse reaction caused by IgG antibodies against platelet factor 4 (PF4). To study VITT, patient samples are crucial but have become a scarce...
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2024-12-01
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| author | Luisa Müller Venkata A. S. Dabbiru Lucy Rutten Rinke Bos Roland Zahn Stefan Handtke Thomas Thiele Marta Palicio Olga Esteban Marta Broto Tom Paul Gordon Andreas Greinacher Jing Jing Wang Linda Schönborn |
| author_facet | Luisa Müller Venkata A. S. Dabbiru Lucy Rutten Rinke Bos Roland Zahn Stefan Handtke Thomas Thiele Marta Palicio Olga Esteban Marta Broto Tom Paul Gordon Andreas Greinacher Jing Jing Wang Linda Schönborn |
| author_sort | Luisa Müller |
| collection | DOAJ |
| description | Background/Objectives: Adenoviral vector-based vaccines against COVID-19 rarely cause vaccine-induced immune thrombocytopenia and thrombosis (VITT), a severe adverse reaction caused by IgG antibodies against platelet factor 4 (PF4). To study VITT, patient samples are crucial but have become a scarce resource. Recombinant antibodies (rAbs) derived from VITT patient characteristic amino acid sequences of anti-PF4 IgG are an alternative to study VITT pathophysiology. Methods: Amino acid sequences of the variable region of immunoglobulin light and heavy chain of anti-PF4 IgG derived from VITT patients were obtained by mass spectrometry sequencing and rAbs were synthetized by reverse-engineering. Six different rAbs were produced: CR23003, CR23004, and CR23005 (from a patient vaccinated with Jcovden, Johnson & Johnson-Janssen (Beerse, Belgium)), CR22046, and CR22050 and CR22066 (from two different patients vaccinated with Vaxzevria, AstraZeneca (Cambridge, UK)). These rAbs were further characterized using anti-PF4 and anti-PF4/heparin IgG ELISAs, rapid anti-PF4 and anti-PF4/polyanion chemiluminescence assays, and PF4-induced platelet activation assay (PIPA) and their capacity to induce procoagulant platelets. Results: rAbs bound to PF4 alone, but not to PF4/polyanion complexes in rapid chemiluminescence assays. Chemiluminescence assays and both anti-PF4 IgG and anti-PF4 IgG/heparin ELISA showed concentration-dependent PF4 binding of all six rAbs, however, with different reactivities among them. PIPA showed a similar, concentration-dependent platelet activation pattern. rAbs varied in their reactivity and the majority of the tested rAbs were able to induce procoagulant platelets. Conclusions: The six rAbs derived from VITT patients reflect VITT-typical binding capacities and the ability to activate platelets. Therefore, these rAbs offer an attractive new option to study VITT pathophysiology. |
| format | Article |
| id | doaj-art-f207a45568694dbe98c7d3ce36e21f59 |
| institution | Kabale University |
| issn | 2076-393X |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
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| series | Vaccines |
| spelling | doaj-art-f207a45568694dbe98c7d3ce36e21f592025-01-24T13:51:38ZengMDPI AGVaccines2076-393X2024-12-01131310.3390/vaccines13010003Recombinant Anti-PF4 Antibodies Derived from Patients with Vaccine-Induced Immune Thrombocytopenia and Thrombosis (VITT) Facilitate Research and Laboratory Diagnosis of VITTLuisa Müller0Venkata A. S. Dabbiru1Lucy Rutten2Rinke Bos3Roland Zahn4Stefan Handtke5Thomas Thiele6Marta Palicio7Olga Esteban8Marta Broto9Tom Paul Gordon10Andreas Greinacher11Jing Jing Wang12Linda Schönborn13Institut für Transfusionsmedizin, Universitätsmedizin Greifswald, 17489 Greifswald, GermanyInstitut für Transfusionsmedizin, Universitätsmedizin Greifswald, 17489 Greifswald, GermanyJanssen Vaccines & Prevention BV, 2333 CN Leiden, The NetherlandsJanssen Vaccines & Prevention BV, 2333 CN Leiden, The NetherlandsJanssen Vaccines & Prevention BV, 2333 CN Leiden, The NetherlandsInstitut für Transfusionsmedizin, Universitätsmedizin Greifswald, 17489 Greifswald, GermanyInstitut für Transfusionsmedizin, Universitätsmedizin Greifswald, 17489 Greifswald, GermanyWerfen, Lliçà d’Amunt, 08186 Barcelona, SpainWerfen, Lliçà d’Amunt, 08186 Barcelona, SpainWerfen, Lliçà d’Amunt, 08186 Barcelona, SpainDepartment of Immunology, College of Medicine and Public Health, Flinders University and SA Pathology, Bedford Park, Adelaide, SA 5042, AustraliaInstitut für Transfusionsmedizin, Universitätsmedizin Greifswald, 17489 Greifswald, GermanyDepartment of Immunology, College of Medicine and Public Health, Flinders University and SA Pathology, Bedford Park, Adelaide, SA 5042, AustraliaInstitut für Transfusionsmedizin, Universitätsmedizin Greifswald, 17489 Greifswald, GermanyBackground/Objectives: Adenoviral vector-based vaccines against COVID-19 rarely cause vaccine-induced immune thrombocytopenia and thrombosis (VITT), a severe adverse reaction caused by IgG antibodies against platelet factor 4 (PF4). To study VITT, patient samples are crucial but have become a scarce resource. Recombinant antibodies (rAbs) derived from VITT patient characteristic amino acid sequences of anti-PF4 IgG are an alternative to study VITT pathophysiology. Methods: Amino acid sequences of the variable region of immunoglobulin light and heavy chain of anti-PF4 IgG derived from VITT patients were obtained by mass spectrometry sequencing and rAbs were synthetized by reverse-engineering. Six different rAbs were produced: CR23003, CR23004, and CR23005 (from a patient vaccinated with Jcovden, Johnson & Johnson-Janssen (Beerse, Belgium)), CR22046, and CR22050 and CR22066 (from two different patients vaccinated with Vaxzevria, AstraZeneca (Cambridge, UK)). These rAbs were further characterized using anti-PF4 and anti-PF4/heparin IgG ELISAs, rapid anti-PF4 and anti-PF4/polyanion chemiluminescence assays, and PF4-induced platelet activation assay (PIPA) and their capacity to induce procoagulant platelets. Results: rAbs bound to PF4 alone, but not to PF4/polyanion complexes in rapid chemiluminescence assays. Chemiluminescence assays and both anti-PF4 IgG and anti-PF4 IgG/heparin ELISA showed concentration-dependent PF4 binding of all six rAbs, however, with different reactivities among them. PIPA showed a similar, concentration-dependent platelet activation pattern. rAbs varied in their reactivity and the majority of the tested rAbs were able to induce procoagulant platelets. Conclusions: The six rAbs derived from VITT patients reflect VITT-typical binding capacities and the ability to activate platelets. Therefore, these rAbs offer an attractive new option to study VITT pathophysiology.https://www.mdpi.com/2076-393X/13/1/3vaccine-induced immune thrombocytopenia and thrombosis (VITT)platelet-factor 4recombinant antibodiesplatelet activation assay |
| spellingShingle | Luisa Müller Venkata A. S. Dabbiru Lucy Rutten Rinke Bos Roland Zahn Stefan Handtke Thomas Thiele Marta Palicio Olga Esteban Marta Broto Tom Paul Gordon Andreas Greinacher Jing Jing Wang Linda Schönborn Recombinant Anti-PF4 Antibodies Derived from Patients with Vaccine-Induced Immune Thrombocytopenia and Thrombosis (VITT) Facilitate Research and Laboratory Diagnosis of VITT Vaccines vaccine-induced immune thrombocytopenia and thrombosis (VITT) platelet-factor 4 recombinant antibodies platelet activation assay |
| title | Recombinant Anti-PF4 Antibodies Derived from Patients with Vaccine-Induced Immune Thrombocytopenia and Thrombosis (VITT) Facilitate Research and Laboratory Diagnosis of VITT |
| title_full | Recombinant Anti-PF4 Antibodies Derived from Patients with Vaccine-Induced Immune Thrombocytopenia and Thrombosis (VITT) Facilitate Research and Laboratory Diagnosis of VITT |
| title_fullStr | Recombinant Anti-PF4 Antibodies Derived from Patients with Vaccine-Induced Immune Thrombocytopenia and Thrombosis (VITT) Facilitate Research and Laboratory Diagnosis of VITT |
| title_full_unstemmed | Recombinant Anti-PF4 Antibodies Derived from Patients with Vaccine-Induced Immune Thrombocytopenia and Thrombosis (VITT) Facilitate Research and Laboratory Diagnosis of VITT |
| title_short | Recombinant Anti-PF4 Antibodies Derived from Patients with Vaccine-Induced Immune Thrombocytopenia and Thrombosis (VITT) Facilitate Research and Laboratory Diagnosis of VITT |
| title_sort | recombinant anti pf4 antibodies derived from patients with vaccine induced immune thrombocytopenia and thrombosis vitt facilitate research and laboratory diagnosis of vitt |
| topic | vaccine-induced immune thrombocytopenia and thrombosis (VITT) platelet-factor 4 recombinant antibodies platelet activation assay |
| url | https://www.mdpi.com/2076-393X/13/1/3 |
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