Primary porcine kupffer cell phagocytosis of human platelets involves the CD18 receptor

Ray K. Chihara, Leela L. Paris, Luz M. Reyes, Richard A. Sidner, Jose L. Estrada, Susan M. Downey, Zheng Yu Wang, A. Joseph Tector, Christopher Burlak

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Background.: Hepatic failure has been treated successfully with clinical extracorporeal perfusions of porcine livers. However, dog-to-pig and pig-to-baboon liver xenotransplant models have resulted in severe bleeding secondary to liver xenograft-induced thrombocytopenia. Kupffer cells (KC) are abundant phagocytic cells in the liver. KC express the CD11b/CD18 receptor, which has been implicated in chilled platelet binding and phagocytosis through interaction with platelet surface proteins and carbohydrates. We sought to identify the role of KC CD18 in liver xenograft-induced thrombocytopenia. Methods.: Primary pig KC were characterized by flow cytometry, immunoblots, and quantitative polymerase chain reaction. Pig KC were used in inhibition assays with fluorescently labeled human platelets. The CD18 receptor was targeted for siRNA knockdown. Results.: Domestic and α1,3-galactosyltransferase double knockout porcine KC cultures were approximately 92% positive for CD18 as detected by quantitative polymerase chain reaction and flow cytometry. Use of CD18 blocking antibodies resulted in reduction of human platelet binding and phagocytosis. Additionally, asialofetuin, not fetuin, inhibited platelet phagocytosis suggesting the involvement of an oligosaccharide-binding site. Furthermore, reduced CD18 expression by siRNA resulted in decreased human platelet binding. Conclusions.: Our data suggest that primary pig KC bind and phagocytose human platelets with involvement of CD18. Further understanding and modification of CD18 expression in pigs may result in a liver xenograft with reduced thrombocytopenic effects, which could be used as a bridge to allogeneic liver transplantation.

Original languageEnglish
Pages (from-to)739-744
Number of pages6
JournalTransplantation
Volume92
Issue number7
DOIs
StatePublished - Oct 15 2011

Fingerprint

Cytophagocytosis
Kupffer Cells
Swine
Blood Platelets
Liver
Phagocytosis
Heterografts
Thrombocytopenia
Small Interfering RNA
Flow Cytometry
Fetuins
Galactosyltransferases
Polymerase Chain Reaction
Blocking Antibodies
Papio
Homologous Transplantation
Liver Failure
Phagocytes
Oligosaccharides
Liver Transplantation

Keywords

  • Kupffer cell
  • Macrophage antigen complex 1 (Mac-1)
  • Phagocytosis
  • thrombocytopenia
  • Xenotransplantation

ASJC Scopus subject areas

  • Transplantation

Cite this

Chihara, R. K., Paris, L. L., Reyes, L. M., Sidner, R. A., Estrada, J. L., Downey, S. M., ... Burlak, C. (2011). Primary porcine kupffer cell phagocytosis of human platelets involves the CD18 receptor. Transplantation, 92(7), 739-744. https://doi.org/10.1097/TP.0b013e31822bc986

Primary porcine kupffer cell phagocytosis of human platelets involves the CD18 receptor. / Chihara, Ray K.; Paris, Leela L.; Reyes, Luz M.; Sidner, Richard A.; Estrada, Jose L.; Downey, Susan M.; Wang, Zheng Yu; Tector, A. Joseph; Burlak, Christopher.

In: Transplantation, Vol. 92, No. 7, 15.10.2011, p. 739-744.

Research output: Contribution to journalArticle

Chihara, RK, Paris, LL, Reyes, LM, Sidner, RA, Estrada, JL, Downey, SM, Wang, ZY, Tector, AJ & Burlak, C 2011, 'Primary porcine kupffer cell phagocytosis of human platelets involves the CD18 receptor', Transplantation, vol. 92, no. 7, pp. 739-744. https://doi.org/10.1097/TP.0b013e31822bc986
Chihara RK, Paris LL, Reyes LM, Sidner RA, Estrada JL, Downey SM et al. Primary porcine kupffer cell phagocytosis of human platelets involves the CD18 receptor. Transplantation. 2011 Oct 15;92(7):739-744. https://doi.org/10.1097/TP.0b013e31822bc986
Chihara, Ray K. ; Paris, Leela L. ; Reyes, Luz M. ; Sidner, Richard A. ; Estrada, Jose L. ; Downey, Susan M. ; Wang, Zheng Yu ; Tector, A. Joseph ; Burlak, Christopher. / Primary porcine kupffer cell phagocytosis of human platelets involves the CD18 receptor. In: Transplantation. 2011 ; Vol. 92, No. 7. pp. 739-744.
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AU - Paris, Leela L.

AU - Reyes, Luz M.

AU - Sidner, Richard A.

AU - Estrada, Jose L.

AU - Downey, Susan M.

AU - Wang, Zheng Yu

AU - Tector, A. Joseph

AU - Burlak, Christopher

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N2 - Background.: Hepatic failure has been treated successfully with clinical extracorporeal perfusions of porcine livers. However, dog-to-pig and pig-to-baboon liver xenotransplant models have resulted in severe bleeding secondary to liver xenograft-induced thrombocytopenia. Kupffer cells (KC) are abundant phagocytic cells in the liver. KC express the CD11b/CD18 receptor, which has been implicated in chilled platelet binding and phagocytosis through interaction with platelet surface proteins and carbohydrates. We sought to identify the role of KC CD18 in liver xenograft-induced thrombocytopenia. Methods.: Primary pig KC were characterized by flow cytometry, immunoblots, and quantitative polymerase chain reaction. Pig KC were used in inhibition assays with fluorescently labeled human platelets. The CD18 receptor was targeted for siRNA knockdown. Results.: Domestic and α1,3-galactosyltransferase double knockout porcine KC cultures were approximately 92% positive for CD18 as detected by quantitative polymerase chain reaction and flow cytometry. Use of CD18 blocking antibodies resulted in reduction of human platelet binding and phagocytosis. Additionally, asialofetuin, not fetuin, inhibited platelet phagocytosis suggesting the involvement of an oligosaccharide-binding site. Furthermore, reduced CD18 expression by siRNA resulted in decreased human platelet binding. Conclusions.: Our data suggest that primary pig KC bind and phagocytose human platelets with involvement of CD18. Further understanding and modification of CD18 expression in pigs may result in a liver xenograft with reduced thrombocytopenic effects, which could be used as a bridge to allogeneic liver transplantation.

AB - Background.: Hepatic failure has been treated successfully with clinical extracorporeal perfusions of porcine livers. However, dog-to-pig and pig-to-baboon liver xenotransplant models have resulted in severe bleeding secondary to liver xenograft-induced thrombocytopenia. Kupffer cells (KC) are abundant phagocytic cells in the liver. KC express the CD11b/CD18 receptor, which has been implicated in chilled platelet binding and phagocytosis through interaction with platelet surface proteins and carbohydrates. We sought to identify the role of KC CD18 in liver xenograft-induced thrombocytopenia. Methods.: Primary pig KC were characterized by flow cytometry, immunoblots, and quantitative polymerase chain reaction. Pig KC were used in inhibition assays with fluorescently labeled human platelets. The CD18 receptor was targeted for siRNA knockdown. Results.: Domestic and α1,3-galactosyltransferase double knockout porcine KC cultures were approximately 92% positive for CD18 as detected by quantitative polymerase chain reaction and flow cytometry. Use of CD18 blocking antibodies resulted in reduction of human platelet binding and phagocytosis. Additionally, asialofetuin, not fetuin, inhibited platelet phagocytosis suggesting the involvement of an oligosaccharide-binding site. Furthermore, reduced CD18 expression by siRNA resulted in decreased human platelet binding. Conclusions.: Our data suggest that primary pig KC bind and phagocytose human platelets with involvement of CD18. Further understanding and modification of CD18 expression in pigs may result in a liver xenograft with reduced thrombocytopenic effects, which could be used as a bridge to allogeneic liver transplantation.

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