Anti–Human Neutrophil Antigen-3a Induced Transfusion-Related Acute Lung Injury in Mice by Direct Disturbance of Lung Endothelial Cells

• Published in: Arteriosclerosis, thrombosis, and vascular biology Vol. 33; no. 11; pp. 2538 - 2548
• Main Authors: Bayat, Behnaz, Tjahjono, Yudy, Sydykov, Akylbek, Werth, Silke, Hippenstiel, Stefan, Weissmann, Nobert, Sachs, Ulrich J., Santoso, Sentot
• Format: Journal Article
• Language: English
• Published: United States American Heart Association, Inc 01.11.2013
• Subjects: Actins - metabolism; Acute Lung Injury - etiology; Acute Lung Injury - immunology; Animals; Antibodies - immunology; Antibodies - pharmacology; Bronchoalveolar Lavage Fluid - cytology; Bronchoalveolar Lavage Fluid - immunology; Capillary Permeability - immunology; Disease Models, Animal; Endothelium, Vascular - immunology; Endothelium, Vascular - metabolism; Humans; Isoantigens - immunology; Lipopolysaccharides - pharmacology; Membrane Glycoproteins - genetics; Membrane Glycoproteins - immunology; Membrane Transport Proteins - immunology; Membrane Transport Proteins - metabolism; Mice; Mice, Knockout; NADPH Oxidase 2; NADPH Oxidases - genetics; Neutrophils - immunology; Respiratory Mucosa - cytology; Respiratory Mucosa - immunology; Transfusion Reaction; endothelium; acute lung injury; reactive oxygen species
• ISSN: 1079-5642; 1524-4636; 1524-4636
• DOI: 10.1161/ATVBAHA.113.301206

OBJECTIVE—Antibodies against human neutrophil antigen-3a (HNA-3a) located on choline transporter-like protein 2 induce severe transfusion-related acute lung injury (TRALI). This study aims to identify the mechanism implicated in anti–HNA-3a-mediated TRALI. APPROACH AND RESULTS—Our analysis shows that anti–HNA-3a recognizes 2 choline transporter-like protein 2 isoforms (P1 and P2) on human microvascular endothelial cells from lung blood vessels but reacts only with the P1 isoform on neutrophils. Direct treatment of HNA-3a–positive endothelial cells with anti–HNA-3a, but not with anti–HNA-3b, leads to reactive oxygen species production, increased albumin influx, and decreased endothelial resistance associated with the formation of actin stress filaments and loosening of junctional vascular endothelium-cadherin. In a novel in vivo mouse model, TRALI was documented by significant increase in lung water content, albumin concentration, and neutrophil numbers in the bronchoalveolar lavage on injection of human anti–HNA-3a in lipopolysaccharides-treated, as well as nontreated mice. Interestingly, although neutrophil depletion alleviated severity of lung injury, it failed to prevent TRALI in this model. Infusion of anti–HNA-3a F(ab′)2 fragments caused moderate TRALI. Finally, mice lacking nicotinamide adenine dinucleotide phosphate oxidase (NOX2) were protected from anti–HNA-3a-mediated TRALI. CONCLUSIONS—These data demonstrate the initiation of endothelial barrier dysfunction in vitro and in vivo by direct binding of anti–HNA-3a on endothelial cells. It seems, however, that the presence of neutrophils aggravates barrier dysfunction. This novel mechanism of TRALI primarily mediated by endothelial cell dysfunction via choline transporter-like protein 2 may help to define new treatment strategies to decrease TRALI-related mortality.