Expression of NeuGc on Pig Corneas and Its Potential Significance in Pig Corneal Xenotransplantation

Whayoung Lee, Yuko Miyagawa, Cassandra Long, Burcin Ekser, Eric Walters, Jagdeece Ramsoondar, David Ayares, A. Joseph Tector, David K C Cooper, Hidetaka Hara

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Purpose: Pigs expressing neither galactose-α1,3-galactose (Gal) nor N-glycolylneuraminic acid (NeuGc) take xenotransplantation one step closer to the clinic. Our aims were (1) to document the lack of NeuGc expression on corneas and aortas and cultured endothelial cells [aortic endothelial cells (AECs); corneal (CECs)] of GTKO/NeuGcKO pigs, and (2) to investigate whether the absence of NeuGc reduced human antibody binding to the tissues and cells. Methods: Wild-type (WT), GTKO, and GTKO/NeuGcKO pigs were used for the study. Human tissues and cultured cells were negative controls. Immunofluorescence staining was performed using anti-Gal and anti-NeuGc antibodies, and human IgM and IgG binding to tissues was determined. Flow cytometric analysis was used to determine Gal and NeuGc expression on cultured CECs and AECs and to measure human IgM/IgG binding to these cells. Results: Both Gal and NeuGc were detected on WT pig corneas and aortas. Although GTKO pigs expressed NeuGc, neither humans nor GTKO/NeuGcKO pigs expressed Gal or NeuGc. Human IgM/IgG binding to corneas and aortas from GTKO and GTKO/NeuGcKO pigs was reduced compared with binding to WT pigs. Human antibody binding to GTKO/NeuGcKO AECs was significantly less than that to GTKO AECs, but there was no significant difference in binding between GTKO and GTKO/NeuGcKO CECs. Conclusions: The absence of NeuGc on GTKO aortic tissue and AECs is associated with reduced human antibody binding, and possibly will provide a better outcome in clinical xenotransplantation using vascularized organs. For clinical corneal xenotransplantation, the absence of NeuGc expression on GTKO/NeuGcKO pig corneas may not prove an advantage over GTKO corneas.

Original languageEnglish (US)
Pages (from-to)105-113
Number of pages9
JournalCornea
Volume35
Issue number1
DOIs
StatePublished - Nov 12 2015

Fingerprint

Heterologous Transplantation
Cornea
Swine
Galactose
Endothelial Cells
Immunoglobulin M
Aorta
Immunoglobulin G
Antibodies
Cultured Cells
Fluorescent Antibody Technique
Anti-Idiotypic Antibodies
Staining and Labeling

Keywords

  • anti-pig
  • antibody
  • cornea
  • immune response
  • N-glycolylneuraminic acid
  • pig
  • porcine
  • xenotransplantation

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Lee, W., Miyagawa, Y., Long, C., Ekser, B., Walters, E., Ramsoondar, J., ... Hara, H. (2015). Expression of NeuGc on Pig Corneas and Its Potential Significance in Pig Corneal Xenotransplantation. Cornea, 35(1), 105-113. https://doi.org/10.1097/ICO.0000000000000635

Expression of NeuGc on Pig Corneas and Its Potential Significance in Pig Corneal Xenotransplantation. / Lee, Whayoung; Miyagawa, Yuko; Long, Cassandra; Ekser, Burcin; Walters, Eric; Ramsoondar, Jagdeece; Ayares, David; Tector, A. Joseph; Cooper, David K C; Hara, Hidetaka.

In: Cornea, Vol. 35, No. 1, 12.11.2015, p. 105-113.

Research output: Contribution to journalArticle

Lee, W, Miyagawa, Y, Long, C, Ekser, B, Walters, E, Ramsoondar, J, Ayares, D, Tector, AJ, Cooper, DKC & Hara, H 2015, 'Expression of NeuGc on Pig Corneas and Its Potential Significance in Pig Corneal Xenotransplantation', Cornea, vol. 35, no. 1, pp. 105-113. https://doi.org/10.1097/ICO.0000000000000635
Lee, Whayoung ; Miyagawa, Yuko ; Long, Cassandra ; Ekser, Burcin ; Walters, Eric ; Ramsoondar, Jagdeece ; Ayares, David ; Tector, A. Joseph ; Cooper, David K C ; Hara, Hidetaka. / Expression of NeuGc on Pig Corneas and Its Potential Significance in Pig Corneal Xenotransplantation. In: Cornea. 2015 ; Vol. 35, No. 1. pp. 105-113.
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AU - Lee, Whayoung

AU - Miyagawa, Yuko

AU - Long, Cassandra

AU - Ekser, Burcin

AU - Walters, Eric

AU - Ramsoondar, Jagdeece

AU - Ayares, David

AU - Tector, A. Joseph

AU - Cooper, David K C

AU - Hara, Hidetaka

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N2 - Purpose: Pigs expressing neither galactose-α1,3-galactose (Gal) nor N-glycolylneuraminic acid (NeuGc) take xenotransplantation one step closer to the clinic. Our aims were (1) to document the lack of NeuGc expression on corneas and aortas and cultured endothelial cells [aortic endothelial cells (AECs); corneal (CECs)] of GTKO/NeuGcKO pigs, and (2) to investigate whether the absence of NeuGc reduced human antibody binding to the tissues and cells. Methods: Wild-type (WT), GTKO, and GTKO/NeuGcKO pigs were used for the study. Human tissues and cultured cells were negative controls. Immunofluorescence staining was performed using anti-Gal and anti-NeuGc antibodies, and human IgM and IgG binding to tissues was determined. Flow cytometric analysis was used to determine Gal and NeuGc expression on cultured CECs and AECs and to measure human IgM/IgG binding to these cells. Results: Both Gal and NeuGc were detected on WT pig corneas and aortas. Although GTKO pigs expressed NeuGc, neither humans nor GTKO/NeuGcKO pigs expressed Gal or NeuGc. Human IgM/IgG binding to corneas and aortas from GTKO and GTKO/NeuGcKO pigs was reduced compared with binding to WT pigs. Human antibody binding to GTKO/NeuGcKO AECs was significantly less than that to GTKO AECs, but there was no significant difference in binding between GTKO and GTKO/NeuGcKO CECs. Conclusions: The absence of NeuGc on GTKO aortic tissue and AECs is associated with reduced human antibody binding, and possibly will provide a better outcome in clinical xenotransplantation using vascularized organs. For clinical corneal xenotransplantation, the absence of NeuGc expression on GTKO/NeuGcKO pig corneas may not prove an advantage over GTKO corneas.

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