J Korean Med Sci.  2009 Dec;24(6):1126-1131. 10.3346/jkms.2009.24.6.1126.

Removal of Alpha-Gal Epitopes from Porcine Aortic Valve and Pericardium using Recombinant Human Alpha Galactosidase A

• Park S
• Kim WH
• Choi SY
• Kim YJ

• KMID: 1783143
• DOI: http://doi.org/10.3346/jkms.2009.24.6.1126

Abstract

It has been reported that the immune response due to alpha-Gal epitopes is an important factor in tissue valve failure. The elimination of the interaction between the natural anti-Gal antibodies and alpha-gal epitopes on the xenografts is a prerequisite to the success of xenografts in humans. Previously, we reported that the green coffee bean alpha-galactosidase could remove all alpha-Gal epitopes from cell surface of porcine aortic valve and pericardial tissue, but it has limitations on cost effectiveness. In this study we wanted to know whether the recently produced recombinant human alpha-galactosidase A has the same effective enzymatic activity as green coffee bean alpha-galactosidase in removing alpha-Gal epitopes from the same tissues. After treating fresh porcine aortic valve and pericardial tissue with recombinant alpha-galactosidase A, each sample was stained with Griffonia simplicifolia type I isolectin B4 indirect immunoperoxidase avidin-biotin technique. We then examined whether the alpha-Gal epitopes were reduced or abolished in each consecutive concentration of recombinant alpha-galactosidase A by comparing the degree of the Griffonia simplicifolia isolectin B4 staining. As a result, the recombinant alpha-galactosidase A could remove cell surface alpha-Gals on porcine aortic valve and pericardial tissue as effectively as green coffee bean alpha-galactosidase.

Keyword

Alpha-Galactosyl Epitope; Alpha-Galactosidase; Bioprosthesis

MeSH Terms

Adolescent
Animals
Aortic Valve/chemistry/cytology/*immunology
Child
Coffea/enzymology
Epitopes/*immunology
Heart Valve Prosthesis Implantation
Humans
Pericardium/chemistry/cytology/*immunology
Recombinant Proteins/genetics/*immunology
Swine
Transplantation, Heterologous/immunology
alpha-Galactosidase/genetics/*immunology

Figure

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  Fig. 1 Aortic valve of a pig (×400): (A) before treatment, (B) after treatment with 1.0 unit/mL of recombinant α-galactosidase A. Many α-Gal epitopes are still present on cell surface (arrow).

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  Fig. 2 Aortic valve of a pig (×400): (A) before treatment, (B) after treatment with 5.0 unit/mL of recombinant α-galactosidase A. α-Gal epitopes are nearly completely removed from cell surface.

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  Fig. 3 Aortic valve of a pig (×400): (A) before treatment, (B) after treatment with 10.0 unit/mL of recombinant α-galactosidase A. No α-Gal epitopes on cell surface.

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  Fig. 4 Pericardium of a pig (×400): (A) before treatment, (B) after treatment with 1.0 unit/mL of recombinant α-galactosidase A. Many α-Gal epitopes are still present on cell surface (arrow).

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  Fig. 5 Pericardium of a pig (×400): (A) before treatment, (B) after treatment with 5.0 unit/mL of recombinant α-galactosidase A. Some α-Gal epitopes are still present on cell surface (arrow).

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  Fig. 6 Pericardium of a pig (×400): (A) before treatment, (B) after treatment with 10.0 unit/mL of recombinant α-galactosidase A. α-Gal epitopes are nearly completely removed from cell surface.

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