ASGR1 expressed by porcine enriched liver sinusoidal endothelial cells mediates human platelet phagocytosis in vitro

Paris LL, Chihara RK, Reyes LM, Sidner RA, Estrada JL, Downey SM, Milgrom DA, Joseph Tector A, Burlak C. ASGR1 expressed by porcine enriched liver sinusoidal endothelial cells mediates human platelet phagocytosis in vitro. Xenotransplantation 2011; 18: 245–251. © 2011 John Wiley & Sons A/S. :  B...

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Published in:Xenotransplantation (Københaven) 2011-07, Vol.18 (4), p.245-251
Main Authors: Paris, Leela L., Chihara, Ray K., Reyes, Luz M., Sidner, Richard A., Estrada, Jose L., Downey, Susan M., Milgrom, Daniel A., Tector, A. Joseph, Burlak, Christopher
Format: Article
Language:English
Subjects:
Animals
Antibodies
Asialofetuin
Asialoglycoprotein Receptor - genetics
Asialoglycoprotein Receptor - metabolism
asialoglycoprotein receptor-1
Blood Platelets - metabolism
Buffers
Cells, Cultured
Confocal microscopy
Dopamine
endothelial cell
Endothelial cells
Endothelial Cells - metabolism
Endothelium
Flow cytometry
Galactose
Glucosamine
Hepatocytes
Humans
Immunohistochemistry
Liver
Liver - cytology
Phagocytosis
Phagocytosis - physiology
Platelet Transfusion
Platelets
Polymerase chain reaction
RNA, Small Interfering - genetics
RNA, Small Interfering - metabolism
siRNA
Swine
Therapeutic applications
Thrombocytopenia
Transplantation, Heterologous
Xenografts
xenotransplantation
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Summary:Paris LL, Chihara RK, Reyes LM, Sidner RA, Estrada JL, Downey SM, Milgrom DA, Joseph Tector A, Burlak C. ASGR1 expressed by porcine enriched liver sinusoidal endothelial cells mediates human platelet phagocytosis in vitro. Xenotransplantation 2011; 18: 245–251. © 2011 John Wiley & Sons A/S. :  Background:  Porcine liver xenografts represent a potential solution to the organ shortage, but thrombocytopenia occurs within minutes to hours after xenotransplantation, preventing clinical application. Recently, it was discovered that porcine liver sinusoidal endothelial cells (LSEC) bind and phagocytose human platelets. We examined the role of ASGR1 in binding and removing human platelets by the pig liver endothelium. Methods:  Primary porcine enriched LSEC (eLSEC) were characterized by flow cytometry, immunoblot, quantitative PCR, and immunohistochemistry using confocal microscopy. Phagocytosis inhibition assays using anti‐ASGR1 and an ASGR1 substrate were performed. ASGR1 was targeted for siRNA knockdown, and ASGR1‐reduced cells were tested for human platelet binding and phagocytosis. Results:  ASGR1 is expressed by eLSEC. Human platelet binding and phagocytosis by porcine eLSEC was inhibited by asialofetuin, but not fetuin, suggesting an interaction with galactose β1‐4 N‐acetyl glucosamine. Anti‐ASGR1 antibodies inhibited human platelet binding in a dose‐dependent manner. Knockdown experiments using siRNA reduced ASGR1 expression in asynchronous primary eLSEC by 40%–80%. There was a 20% reduction in translated protein significantly correlated with a 21% decrease in human platelet binding. Conclusions:  ASGR1 on porcine eLSEC mediates phagocytosis of xenogeneic platelets.
ISSN:0908-665X
1399-3089
DOI:10.1111/j.1399-3089.2011.00639.x