Al-Mohannadi Anjud, Yahia Reem Mohammed, Bibawi Hani, Lachica Che-Ann, Ahmed Watfa, Pavlovski Igor, Gentilcore Giusy, Elgaali Elkhansa Elbukhari, Ejaz Anila, Ahmed Areeg, Elanbari Mohammed, Awada Zainab, Al-Kubaisi Mohammed J, Elnaggar Muhammad, Saleh Ayman, Cugno Chiara, Deola Sara
Research Department, Sidra Medicine, Doha P.O. Box 26999, Qatar.
College of Health and Life Sciences, Hamad Bin Khalifa University, Doha P.O. Box 34110, Qatar.
Cells. 2025 Jan 8;14(2):73. doi: 10.3390/cells14020073.
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.
甲型血友病（HA）与凝血因子VIII（FVIII）凝血功能不全或无活性相关，可导致出血过多。相反，FVIII水平升高与易栓症、血栓形成、心肌梗死和中风相关。活性FVIII（aFVIII）利用其C2结构域与血细胞细胞膜结合，从而发挥其凝血功能。我们开发了一种可靠的流式细胞术（FC）方法用于FVIII检测，该方法可用于评估不同凝血/临床状态下白细胞表面结合的FVIII；我们分析了49名儿科受试者，包括HA患者、其他凝血病患者、静脉血栓形成患者和凝血功能正常者。有趣的是，总白细胞表面FVIII在血栓形成、正常和低凝血状态下呈下降趋势。正如预期的那样，与血栓形成患者相比，HA患者的白细胞表面FVIII水平显著降低。然而，血浆中FVIII的循环水平与白细胞结合的FVIII水平之间未观察到显著相关性，这表明FVIII表面结合的差异与循环中FVIII的可用性不成正比，并提示存在一种特定的结合机制来控制FVIII与白细胞之间的相互作用。有趣的是，在分析不同的血液亚群时，我们观察到与HA患者、健康对照和其他凝血病患者相比，血栓形成患者的经典单核细胞表面FVIII水平显著升高。我们的研究强调了我们的FC平台在评估不同凝血状态下白细胞膜上FVIII丰度方面的可靠性。单核细胞，特别是在血栓形成的情况下，在其表面表现出FVIII的活性结合，这表明其在血栓形成的病理生理学中可能发挥作用，需要进一步研究。
