Flow Cytometry Evaluation of Blood-Cell-Bound Surface FVIII in Hemophilia A and Thrombosis
Hemophilia A (HA) is associated with FVIII coagulation insufficiency or inactivity leading to excessive bleeding. Elevated FVIII, on the contrary, is associated with thrombophilia, thrombosis, myocardial infarctions, and stroke. Active FVIII (aFVIII) uses its C2 domain to bind to blood cells’ membra...
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2025-01-01
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| author | Anjud Al-Mohannadi Reem Mohammed Yahia Hani Bibawi Che-Ann Lachica Watfa Ahmed Igor Pavlovski Giusy Gentilcore Elkhansa Elbukhari Elgaali Anila Ejaz Areeg Ahmed Mohammed Elanbari Zainab Awada Mohammed J. Al-Kubaisi Muhammad Elnaggar Ayman Saleh Chiara Cugno Sara Deola |
| author_facet | Anjud Al-Mohannadi Reem Mohammed Yahia Hani Bibawi Che-Ann Lachica Watfa Ahmed Igor Pavlovski Giusy Gentilcore Elkhansa Elbukhari Elgaali Anila Ejaz Areeg Ahmed Mohammed Elanbari Zainab Awada Mohammed J. Al-Kubaisi Muhammad Elnaggar Ayman Saleh Chiara Cugno Sara Deola |
| author_sort | Anjud Al-Mohannadi |
| collection | DOAJ |
| description | Hemophilia A (HA) is associated with FVIII coagulation insufficiency or inactivity leading to excessive bleeding. Elevated FVIII, on the contrary, is associated with thrombophilia, thrombosis, myocardial infarctions, and stroke. Active FVIII (aFVIII) uses its C2 domain to bind to blood cells’ membranes, consequently carrying out its coagulative function. We developed a reliable flow cytometry (FC) method for FVIII detection that can be utilized for assessing surface-bound FVIII on leukocytes in different coagulation/clinical states; we analyzed 49 pediatric subjects, encompassing patients with HA, other coagulopathies, venous thrombosis, and normal coagulation. Interestingly, the total leukocyte surface FVIII showed a declining trend across thrombosis, normal, and hypo-coagulation states. As expected, the leukocytes of HA patients displayed significantly lower levels of cellular-surface FVIII in comparison to patients with thrombosis. However, no significant correlation was observed between circulating levels of FVIII in plasma and the levels of FVIII bound to leukocytes, indicating that the differences in FVIII surface binding are not directly proportional to the availability of FVIII in the circulation and suggesting a specific binding mechanism governing the interaction between FVIII and leukocytes. Intriguingly, when analyzing the distinct blood subpopulations, we observed that surface FVIII levels were significantly elevated in classical monocytes of thrombosis patients compared to HA patients, healthy controls, and patients with other coagulopathies. Our study highlights the reliability of our FC platform in assessing FVIII abundance on leukocytes’ membranes across coagulation states. Monocytes, particularly in cases of thrombosis, exhibit active binding of FVIII on their surface, suggesting a potential role in the pathophysiology of thrombosis that requires further investigation. |
| format | Article |
| id | doaj-art-d432db9a32be4d46a6da27bcefcbbb72 |
| institution | Kabale University |
| issn | 2073-4409 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
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| spelling | doaj-art-d432db9a32be4d46a6da27bcefcbbb722025-01-24T13:26:34ZengMDPI AGCells2073-44092025-01-011427310.3390/cells14020073Flow Cytometry Evaluation of Blood-Cell-Bound Surface FVIII in Hemophilia A and ThrombosisAnjud Al-Mohannadi0Reem Mohammed Yahia1Hani Bibawi2Che-Ann Lachica3Watfa Ahmed4Igor Pavlovski5Giusy Gentilcore6Elkhansa Elbukhari Elgaali7Anila Ejaz8Areeg Ahmed9Mohammed Elanbari10Zainab Awada11Mohammed J. Al-Kubaisi12Muhammad Elnaggar13Ayman Saleh14Chiara Cugno15Sara Deola16Research Department, Sidra Medicine, Doha P.O. Box 26999, QatarResearch Department, Sidra Medicine, Doha P.O. Box 26999, QatarPathology Department, Sidra Medicine, Doha P.O. Box 26999, QatarResearch Department, Sidra Medicine, Doha P.O. Box 26999, QatarResearch Department, Sidra Medicine, Doha P.O. Box 26999, QatarResearch Department, Sidra Medicine, Doha P.O. Box 26999, QatarResearch Department, Sidra Medicine, Doha P.O. Box 26999, QatarHematology-Oncology Clinic, Sidra Medicine, Doha P.O. Box 26999, QatarHematology-Oncology Clinic, Sidra Medicine, Doha P.O. Box 26999, QatarHematology-Oncology Clinic, Sidra Medicine, Doha P.O. Box 26999, QatarResearch Department, Sidra Medicine, Doha P.O. Box 26999, QatarResearch Department, Sidra Medicine, Doha P.O. Box 26999, QatarResearch Department, Sidra Medicine, Doha P.O. Box 26999, QatarResearch Department, Sidra Medicine, Doha P.O. Box 26999, QatarHematology-Oncology Clinic, Sidra Medicine, Doha P.O. Box 26999, QatarResearch Department, Sidra Medicine, Doha P.O. Box 26999, QatarResearch Department, Sidra Medicine, Doha P.O. Box 26999, QatarHemophilia A (HA) is associated with FVIII coagulation insufficiency or inactivity leading to excessive bleeding. Elevated FVIII, on the contrary, is associated with thrombophilia, thrombosis, myocardial infarctions, and stroke. Active FVIII (aFVIII) uses its C2 domain to bind to blood cells’ membranes, consequently carrying out its coagulative function. We developed a reliable flow cytometry (FC) method for FVIII detection that can be utilized for assessing surface-bound FVIII on leukocytes in different coagulation/clinical states; we analyzed 49 pediatric subjects, encompassing patients with HA, other coagulopathies, venous thrombosis, and normal coagulation. Interestingly, the total leukocyte surface FVIII showed a declining trend across thrombosis, normal, and hypo-coagulation states. As expected, the leukocytes of HA patients displayed significantly lower levels of cellular-surface FVIII in comparison to patients with thrombosis. However, no significant correlation was observed between circulating levels of FVIII in plasma and the levels of FVIII bound to leukocytes, indicating that the differences in FVIII surface binding are not directly proportional to the availability of FVIII in the circulation and suggesting a specific binding mechanism governing the interaction between FVIII and leukocytes. Intriguingly, when analyzing the distinct blood subpopulations, we observed that surface FVIII levels were significantly elevated in classical monocytes of thrombosis patients compared to HA patients, healthy controls, and patients with other coagulopathies. Our study highlights the reliability of our FC platform in assessing FVIII abundance on leukocytes’ membranes across coagulation states. Monocytes, particularly in cases of thrombosis, exhibit active binding of FVIII on their surface, suggesting a potential role in the pathophysiology of thrombosis that requires further investigation.https://www.mdpi.com/2073-4409/14/2/73flow cytometryfactor VIIIhemophilia Aleukocytesmonocytescoagulation |
| spellingShingle | Anjud Al-Mohannadi Reem Mohammed Yahia Hani Bibawi Che-Ann Lachica Watfa Ahmed Igor Pavlovski Giusy Gentilcore Elkhansa Elbukhari Elgaali Anila Ejaz Areeg Ahmed Mohammed Elanbari Zainab Awada Mohammed J. Al-Kubaisi Muhammad Elnaggar Ayman Saleh Chiara Cugno Sara Deola Flow Cytometry Evaluation of Blood-Cell-Bound Surface FVIII in Hemophilia A and Thrombosis Cells flow cytometry factor VIII hemophilia A leukocytes monocytes coagulation |
| title | Flow Cytometry Evaluation of Blood-Cell-Bound Surface FVIII in Hemophilia A and Thrombosis |
| title_full | Flow Cytometry Evaluation of Blood-Cell-Bound Surface FVIII in Hemophilia A and Thrombosis |
| title_fullStr | Flow Cytometry Evaluation of Blood-Cell-Bound Surface FVIII in Hemophilia A and Thrombosis |
| title_full_unstemmed | Flow Cytometry Evaluation of Blood-Cell-Bound Surface FVIII in Hemophilia A and Thrombosis |
| title_short | Flow Cytometry Evaluation of Blood-Cell-Bound Surface FVIII in Hemophilia A and Thrombosis |
| title_sort | flow cytometry evaluation of blood cell bound surface fviii in hemophilia a and thrombosis |
| topic | flow cytometry factor VIII hemophilia A leukocytes monocytes coagulation |
| url | https://www.mdpi.com/2073-4409/14/2/73 |
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