Fibronectin from alpha 1,3-galactosyltransferase knockout pigs is a xenoantigen

Ray K. Chihara, Andrew J. Lutz, Leela L. Paris, Zheng Yu Wang, Richard A. Sidner, Alex T. Heyrman, Susan M. Downey, Christopher Burlak, A. Joseph Tector

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Background: Antibody-mediated rejection continues to be an obstacle for xenotransplantation despite development of α1,3-galactosyltransferase knockout (GTKO) pigs. Fibronectin (Fn) from GTKO pigs was identified as a xenoantigen in baboons. N-glycolylneuraminic acid (Neu5Gc), similar to galactose α1,3-galactose, is an antigenic carbohydrate found in pigs. We evaluated human antibody reactivity and performed initial antigenic epitope characterization of Fn from GTKO pigs. Materials and methods: GTKO pig aortic endothelial cells (AEC) were isolated and assessed for antibody-mediated complement-dependent cytotoxicity (CDC). Human and GTKO pig Fn were purified and analyzed using immunoblots. GTKO pig and human AEC absorbed human sera were assessed for CDC and anti-GTKO pig Fn antibodies. GTKO pig proteins were assessed for Neu5Gc. Immunoaffinity-purified human IgG anti-GTKO pig (hIgG-GTKOp) Fn using a GTKO pig Fn column were evaluated for cross-reactivity with other proteins. Results: GTKO pig AEC had greater human antibody binding, complement deposition and CDC compared with allogeneic human AEC. Human sera absorbed with GTKO pig AEC resulted in diminished anti-GTKO pig Fn antibody. Neu5Gc was identified on GTKO pig Fn and other proteins. The hIgG-GTKOp Fn cross-reacted with multiple GTKO pig proteins and was enriched with anti-Neu5Gc antibody. Conclusions: Removal of antigenic epitopes from GTKO pig AEC would improve xenograft compatibility. GTKO pig Fn has antigenic epitopes, one identified as Neu5Gc, which may be responsible for pathology and cross-reactivity of hIgG-GTKOp Fn. Genetic knockout of Neu5Gc appears necessary to address significance and identification of non-Neu5Gc GTKO pig Fn antigenic epitopes.

Original languageEnglish
Pages (from-to)1123-1133
Number of pages11
JournalJournal of Surgical Research
Volume184
Issue number2
DOIs
StatePublished - Oct 2013

Fingerprint

Heterophile Antigens
Galactosyltransferases
Fibronectins
Swine
Endothelial Cells
Antibodies
Epitopes
N-acetyllactosaminide alpha-1,3-galactosyltransferase
Galactose
Proteins

Keywords

  • Antibody-mediated rejection
  • Fibronectin
  • N-glycolylneuraminic acid
  • Neu5Gc
  • Xenoantigen
  • Xenotransplantation

ASJC Scopus subject areas

  • Surgery

Cite this

Chihara, R. K., Lutz, A. J., Paris, L. L., Wang, Z. Y., Sidner, R. A., Heyrman, A. T., ... Tector, A. J. (2013). Fibronectin from alpha 1,3-galactosyltransferase knockout pigs is a xenoantigen. Journal of Surgical Research, 184(2), 1123-1133. https://doi.org/10.1016/j.jss.2013.04.012

Fibronectin from alpha 1,3-galactosyltransferase knockout pigs is a xenoantigen. / Chihara, Ray K.; Lutz, Andrew J.; Paris, Leela L.; Wang, Zheng Yu; Sidner, Richard A.; Heyrman, Alex T.; Downey, Susan M.; Burlak, Christopher; Tector, A. Joseph.

In: Journal of Surgical Research, Vol. 184, No. 2, 10.2013, p. 1123-1133.

Research output: Contribution to journalArticle

Chihara, RK, Lutz, AJ, Paris, LL, Wang, ZY, Sidner, RA, Heyrman, AT, Downey, SM, Burlak, C & Tector, AJ 2013, 'Fibronectin from alpha 1,3-galactosyltransferase knockout pigs is a xenoantigen', Journal of Surgical Research, vol. 184, no. 2, pp. 1123-1133. https://doi.org/10.1016/j.jss.2013.04.012
Chihara RK, Lutz AJ, Paris LL, Wang ZY, Sidner RA, Heyrman AT et al. Fibronectin from alpha 1,3-galactosyltransferase knockout pigs is a xenoantigen. Journal of Surgical Research. 2013 Oct;184(2):1123-1133. https://doi.org/10.1016/j.jss.2013.04.012
Chihara, Ray K. ; Lutz, Andrew J. ; Paris, Leela L. ; Wang, Zheng Yu ; Sidner, Richard A. ; Heyrman, Alex T. ; Downey, Susan M. ; Burlak, Christopher ; Tector, A. Joseph. / Fibronectin from alpha 1,3-galactosyltransferase knockout pigs is a xenoantigen. In: Journal of Surgical Research. 2013 ; Vol. 184, No. 2. pp. 1123-1133.
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abstract = "Background: Antibody-mediated rejection continues to be an obstacle for xenotransplantation despite development of α1,3-galactosyltransferase knockout (GTKO) pigs. Fibronectin (Fn) from GTKO pigs was identified as a xenoantigen in baboons. N-glycolylneuraminic acid (Neu5Gc), similar to galactose α1,3-galactose, is an antigenic carbohydrate found in pigs. We evaluated human antibody reactivity and performed initial antigenic epitope characterization of Fn from GTKO pigs. Materials and methods: GTKO pig aortic endothelial cells (AEC) were isolated and assessed for antibody-mediated complement-dependent cytotoxicity (CDC). Human and GTKO pig Fn were purified and analyzed using immunoblots. GTKO pig and human AEC absorbed human sera were assessed for CDC and anti-GTKO pig Fn antibodies. GTKO pig proteins were assessed for Neu5Gc. Immunoaffinity-purified human IgG anti-GTKO pig (hIgG-GTKOp) Fn using a GTKO pig Fn column were evaluated for cross-reactivity with other proteins. Results: GTKO pig AEC had greater human antibody binding, complement deposition and CDC compared with allogeneic human AEC. Human sera absorbed with GTKO pig AEC resulted in diminished anti-GTKO pig Fn antibody. Neu5Gc was identified on GTKO pig Fn and other proteins. The hIgG-GTKOp Fn cross-reacted with multiple GTKO pig proteins and was enriched with anti-Neu5Gc antibody. Conclusions: Removal of antigenic epitopes from GTKO pig AEC would improve xenograft compatibility. GTKO pig Fn has antigenic epitopes, one identified as Neu5Gc, which may be responsible for pathology and cross-reactivity of hIgG-GTKOp Fn. Genetic knockout of Neu5Gc appears necessary to address significance and identification of non-Neu5Gc GTKO pig Fn antigenic epitopes.",
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T1 - Fibronectin from alpha 1,3-galactosyltransferase knockout pigs is a xenoantigen

AU - Chihara, Ray K.

AU - Lutz, Andrew J.

AU - Paris, Leela L.

AU - Wang, Zheng Yu

AU - Sidner, Richard A.

AU - Heyrman, Alex T.

AU - Downey, Susan M.

AU - Burlak, Christopher

AU - Tector, A. Joseph

PY - 2013/10

Y1 - 2013/10

N2 - Background: Antibody-mediated rejection continues to be an obstacle for xenotransplantation despite development of α1,3-galactosyltransferase knockout (GTKO) pigs. Fibronectin (Fn) from GTKO pigs was identified as a xenoantigen in baboons. N-glycolylneuraminic acid (Neu5Gc), similar to galactose α1,3-galactose, is an antigenic carbohydrate found in pigs. We evaluated human antibody reactivity and performed initial antigenic epitope characterization of Fn from GTKO pigs. Materials and methods: GTKO pig aortic endothelial cells (AEC) were isolated and assessed for antibody-mediated complement-dependent cytotoxicity (CDC). Human and GTKO pig Fn were purified and analyzed using immunoblots. GTKO pig and human AEC absorbed human sera were assessed for CDC and anti-GTKO pig Fn antibodies. GTKO pig proteins were assessed for Neu5Gc. Immunoaffinity-purified human IgG anti-GTKO pig (hIgG-GTKOp) Fn using a GTKO pig Fn column were evaluated for cross-reactivity with other proteins. Results: GTKO pig AEC had greater human antibody binding, complement deposition and CDC compared with allogeneic human AEC. Human sera absorbed with GTKO pig AEC resulted in diminished anti-GTKO pig Fn antibody. Neu5Gc was identified on GTKO pig Fn and other proteins. The hIgG-GTKOp Fn cross-reacted with multiple GTKO pig proteins and was enriched with anti-Neu5Gc antibody. Conclusions: Removal of antigenic epitopes from GTKO pig AEC would improve xenograft compatibility. GTKO pig Fn has antigenic epitopes, one identified as Neu5Gc, which may be responsible for pathology and cross-reactivity of hIgG-GTKOp Fn. Genetic knockout of Neu5Gc appears necessary to address significance and identification of non-Neu5Gc GTKO pig Fn antigenic epitopes.

AB - Background: Antibody-mediated rejection continues to be an obstacle for xenotransplantation despite development of α1,3-galactosyltransferase knockout (GTKO) pigs. Fibronectin (Fn) from GTKO pigs was identified as a xenoantigen in baboons. N-glycolylneuraminic acid (Neu5Gc), similar to galactose α1,3-galactose, is an antigenic carbohydrate found in pigs. We evaluated human antibody reactivity and performed initial antigenic epitope characterization of Fn from GTKO pigs. Materials and methods: GTKO pig aortic endothelial cells (AEC) were isolated and assessed for antibody-mediated complement-dependent cytotoxicity (CDC). Human and GTKO pig Fn were purified and analyzed using immunoblots. GTKO pig and human AEC absorbed human sera were assessed for CDC and anti-GTKO pig Fn antibodies. GTKO pig proteins were assessed for Neu5Gc. Immunoaffinity-purified human IgG anti-GTKO pig (hIgG-GTKOp) Fn using a GTKO pig Fn column were evaluated for cross-reactivity with other proteins. Results: GTKO pig AEC had greater human antibody binding, complement deposition and CDC compared with allogeneic human AEC. Human sera absorbed with GTKO pig AEC resulted in diminished anti-GTKO pig Fn antibody. Neu5Gc was identified on GTKO pig Fn and other proteins. The hIgG-GTKOp Fn cross-reacted with multiple GTKO pig proteins and was enriched with anti-Neu5Gc antibody. Conclusions: Removal of antigenic epitopes from GTKO pig AEC would improve xenograft compatibility. GTKO pig Fn has antigenic epitopes, one identified as Neu5Gc, which may be responsible for pathology and cross-reactivity of hIgG-GTKOp Fn. Genetic knockout of Neu5Gc appears necessary to address significance and identification of non-Neu5Gc GTKO pig Fn antigenic epitopes.

KW - Antibody-mediated rejection

KW - Fibronectin

KW - N-glycolylneuraminic acid

KW - Neu5Gc

KW - Xenoantigen

KW - Xenotransplantation

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