McRae Hannah L, Millar Michelle Warren, Slavin Spencer A, Blumberg Neil, Rahman Arshad, Refaai Majed A
Department of Pathology and Laboratory Medicine, Transfusion Medicine Division, University of Rochester Medical Center, Rochester, NY 14642, USA.
Department of Pediatrics (Neonatology), Lung Biology and Disease Program, University of Rochester Medical Center, Rochester, NY 14642, USA.
Biomedicines. 2021 Dec 7;9(12):1851. doi: 10.3390/biomedicines9121851.
ABO immune complexes (ABO-IC) formed by ABO-incompatible antigen-antibody interaction are associated with hemolysis and platelet destruction in patients transfused with ABO-nonidentical blood products. However, the effects of ABO-IC on endothelial cells (EC) are unclear. ABO-IC were formed in vitro from normal donor-derived plasma and serum. Human pulmonary artery EC (HPAEC) were cultured and treated with media, ABO-identical and -non-identical plasma, and ABO-IC. EC barrier integrity was evaluated using transendothelial electrical resistance (TEER), scanning electron microscopy (SEM), vascular endothelial (VE)-cadherin and phalloidin staining, and Rho-associated Kinase (ROCK) inhibitor treatment. TEER revealed significant/irreversible barrier disruption within 1-2 h of exposure to ABO non-identical plasma and ABO-IC; this occurred independently of EC ABO type. Treatment with ABO-IC resulted in decreased VE-cadherin staining and increased phalloidin staining in a time-dependent manner, suggesting that the resultant increased EC barrier permeability is secondary to actin stress fiber formation and loss of cell surface VE-cadherin. Inhibition of ROCK was effective in protecting against IC-induced barrier disruption even two hours after ABO-IC exposure. ABO-IC causes increased EC barrier permeability by decreasing cell surface VE-cadherin and promoting stress fiber formation, which is preventable by inhibiting ROCK activation to protect against EC contraction and gap formation.
由ABO血型不相容的抗原-抗体相互作用形成的ABO免疫复合物（ABO-IC）与输注ABO血型不匹配血液制品的患者的溶血和血小板破坏有关。然而，ABO-IC对内皮细胞（EC）的影响尚不清楚。ABO-IC在体外由正常供体来源的血浆和血清形成。培养人肺动脉内皮细胞（HPAEC），并用培养基、ABO血型相同和不相同的血浆以及ABO-IC进行处理。使用跨内皮电阻（TEER）、扫描电子显微镜（SEM）、血管内皮（VE）-钙黏蛋白和鬼笔环肽染色以及Rho相关激酶（ROCK）抑制剂处理来评估内皮细胞屏障的完整性。TEER显示，在暴露于ABO血型不相同的血浆和ABO-IC后1-2小时内，内皮细胞屏障出现显著/不可逆的破坏；这种破坏与内皮细胞的ABO血型无关。用ABO-IC处理导致VE-钙黏蛋白染色减少，并使鬼笔环肽染色呈时间依赖性增加，这表明由此导致的内皮细胞屏障通透性增加是肌动蛋白应力纤维形成和细胞表面VE-钙黏蛋白丢失的结果。即使在暴露于ABO-IC两小时后，抑制ROCK也能有效防止IC诱导的屏障破坏。ABO-IC通过减少细胞表面VE-钙黏蛋白并促进应力纤维形成，导致内皮细胞屏障通透性增加，通过抑制ROCK激活来防止内皮细胞收缩和间隙形成可预防这种情况。
