Essential Role of Rho-Associated Kinase in ABO Immune Complex-Mediated Endothelial Barrier Disruption

• Published in: Biomedicines Vol. 9; no. 12; p. 1851
• Main Authors: McRae, Hannah L, Millar, Michelle Warren, Slavin, Spencer A, Blumberg, Neil, Rahman, Arshad, Refaai, Majed A
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 07.12.2021; MDPI
• Subjects: ABO blood groups; ABO system; Actin; Antigen-antibody complexes; Antigens; Blood; Blood groups; Blood platelets; Blood transfusions; Cadherins; Cell surface; Electrical resistivity; Endothelial cells; Endothelium; Enzyme inhibitors; Experiments; immune complexes; Medical research; Mortality; Pathophysiology; Permeability; Phalloidin; Plasma; Proteins; Pulmonary arteries; Pulmonary artery; Respiratory distress syndrome; Rho-associated kinase; Scanning electron microscopy; transfusion; vascular biology; United States > US; Germany; endothelial cells; transfusion; vascular biology; ABO blood groups; immune complexes
• ISSN: 2227-9059; 2227-9059
• 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.