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Presenter: Leela, Paris, Indianapolis, United States
Authors: Leela Paris1, Ray Chihara1, Richard Sidner1, Luz Reyes1, Jose Estrada1, Daniel Milgrom1, Chris Burlak1, A. Joseph Tector1
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ASGR1 on porcine liver sinusoidal endothelial cells (LSEC) is involved in xenogeneic phagocytosis of human platelets in vitro
Leela Paris, Ray Chihara, Richard Sidner, Luz Reyes, Jose Estrada, Daniel Milgrom, Chris Burlak, A. Joseph Tector
Department of Surgery, Indiana University Medical School, Indianapolis, IN, United States
Pig to baboon liver transplantation results in severe thrombocytopenia with resultant mortality in less than 10 days. Extracorporeal porcine liver perfusion with human blood also results in platelet disappearance. Human platelets are phagocytosed by liver sinusoidal endothelial cells (LSEC) and Kupffer Cells during ex vivo perfusions of porcine livers with human platelets. The asialoglycoprotein receptor 1 (ASGR1) has been implicated in phagocytosis of platelets by Kupffer cells and hepatocytes following platelet cold storage or during infection with S. pneumoniae. Expression of ASGR1 was examined on liver cells including endothelial cells and Kupffer cells using immunocytochemistry, immunohistochemistry, PCR, immunoblot analysis, and flow cytometry. ASGR1 was observed on CD31 positive porcine liver endothelial cells. Domestic and a1,3-galactosyltransferase knockout (GTKO) porcine LSEC had 34.9-16.5% decreased phagocytosis of human platelets following incubation with ASGR1 antibodies in vitro. Targeted ASGR1 knockdown in primary domestic and GTKO porcine LSEC yielded a 20% inhibition of human platelet uptake. A decrease in ASGR1 expression over time was directly correlated with diminished human platelet phagocytosis by porcine LSEC. ASGR1 on porcine LSEC plays a role in phagocytosis of human platelets in vitro.
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