In vivo gene transfer of endo-β-galactosidase C removes αGal antigen on erythrocytes and endothelial cells of the organs

Yusuke Miki, Shoichi Maruyama, Da Ge Liu, Takaaki Kobayashi, Fumihiko Sato, Hideaki Shimizu, Sawako Kato, Waichi Sato, Yoshiki Morita, Yukio Yuzawa, Takashi Muramatsu, Seiichi Matsuo

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Background: The presence of Galα1-3Galβ1-4GlcNAc (αGal) in pigs is a formidable barrier for pig-to-primate xenotransplantation. We have reported that administration of recombinant endo-β-galactosidase C (EndoGalC) removes αGal on porcine erythrocytes and kidneys. The present study examined the effects of EndoGalC gene therapy on αGal suppression. Methods: Naked plasmid DNA encoding Igκ-EndoGalC was given to rats by rapid tail vein injection. The expression of αGal in the heart and kidney were studied by lectin staining followed by computer-assisted quantitative analyses. αGal expression on erythrocytes was determined by flow cytometric analyses. Enzymatic activity of EndoGalC in the serum was also determined by evaluating the capacity of EndoGalC in removing αGal epitopes. Elimination of αGal was further studied by injecting antibodies against αGal to rats 2 days after the gene transfer. Results: Administration of 1 mg of Igκ-EndoGalC/pCAGGS plasmid eliminated αGal from the vascular endothelium of the heart and kidney on day 1 and day 2. Between days 4 and 7, αGal started to reappear, but remained suppressed. No serious adverse effect was observed in rats treated with EndoGalC. Flow cytometric analyses showed that Endo-GalC digested 97% of αGal on erythrocytes when measured 4 days after the gene transfer. Enzymatic activity of EndoGalC in the serum peaked on day 1, and significant levels were still observed on day 7. When antibodies against αGal were given, rats treated with EndoGalC showed no change, while all control rats died within 40 min. Conclusions: The present study demonstrates the potential of an EndoGalC gene transfer, using a hydrodynamics-based delivery system, in eliminating αGal from endothelial cells in vivo. The results also ensured that EndoGalC is not harmful suggesting that the production of pigs overexpressing EndoGalC would be a reasonable alternative to pigs deficient in α1,3galactosyltransferase.

Original languageEnglish
Pages (from-to)444-451
Number of pages8
JournalXenotransplantation
Volume11
Issue number5
DOIs
Publication statusPublished - 01-09-2004
Externally publishedYes

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Galactosidases
Endothelial Cells
Erythrocytes
Antigens
Genes
Swine
Kidney
Plasmids
Heterologous Transplantation
Antibodies
Vascular Endothelium
Hydrodynamics
Serum
Lectins
Genetic Therapy
Primates
Tail

All Science Journal Classification (ASJC) codes

  • Immunology
  • Transplantation

Cite this

Miki, Yusuke ; Maruyama, Shoichi ; Liu, Da Ge ; Kobayashi, Takaaki ; Sato, Fumihiko ; Shimizu, Hideaki ; Kato, Sawako ; Sato, Waichi ; Morita, Yoshiki ; Yuzawa, Yukio ; Muramatsu, Takashi ; Matsuo, Seiichi. / In vivo gene transfer of endo-β-galactosidase C removes αGal antigen on erythrocytes and endothelial cells of the organs. In: Xenotransplantation. 2004 ; Vol. 11, No. 5. pp. 444-451.
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title = "In vivo gene transfer of endo-β-galactosidase C removes αGal antigen on erythrocytes and endothelial cells of the organs",
abstract = "Background: The presence of Galα1-3Galβ1-4GlcNAc (αGal) in pigs is a formidable barrier for pig-to-primate xenotransplantation. We have reported that administration of recombinant endo-β-galactosidase C (EndoGalC) removes αGal on porcine erythrocytes and kidneys. The present study examined the effects of EndoGalC gene therapy on αGal suppression. Methods: Naked plasmid DNA encoding Igκ-EndoGalC was given to rats by rapid tail vein injection. The expression of αGal in the heart and kidney were studied by lectin staining followed by computer-assisted quantitative analyses. αGal expression on erythrocytes was determined by flow cytometric analyses. Enzymatic activity of EndoGalC in the serum was also determined by evaluating the capacity of EndoGalC in removing αGal epitopes. Elimination of αGal was further studied by injecting antibodies against αGal to rats 2 days after the gene transfer. Results: Administration of 1 mg of Igκ-EndoGalC/pCAGGS plasmid eliminated αGal from the vascular endothelium of the heart and kidney on day 1 and day 2. Between days 4 and 7, αGal started to reappear, but remained suppressed. No serious adverse effect was observed in rats treated with EndoGalC. Flow cytometric analyses showed that Endo-GalC digested 97{\%} of αGal on erythrocytes when measured 4 days after the gene transfer. Enzymatic activity of EndoGalC in the serum peaked on day 1, and significant levels were still observed on day 7. When antibodies against αGal were given, rats treated with EndoGalC showed no change, while all control rats died within 40 min. Conclusions: The present study demonstrates the potential of an EndoGalC gene transfer, using a hydrodynamics-based delivery system, in eliminating αGal from endothelial cells in vivo. The results also ensured that EndoGalC is not harmful suggesting that the production of pigs overexpressing EndoGalC would be a reasonable alternative to pigs deficient in α1,3galactosyltransferase.",
author = "Yusuke Miki and Shoichi Maruyama and Liu, {Da Ge} and Takaaki Kobayashi and Fumihiko Sato and Hideaki Shimizu and Sawako Kato and Waichi Sato and Yoshiki Morita and Yukio Yuzawa and Takashi Muramatsu and Seiichi Matsuo",
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Miki, Y, Maruyama, S, Liu, DG, Kobayashi, T, Sato, F, Shimizu, H, Kato, S, Sato, W, Morita, Y, Yuzawa, Y, Muramatsu, T & Matsuo, S 2004, 'In vivo gene transfer of endo-β-galactosidase C removes αGal antigen on erythrocytes and endothelial cells of the organs', Xenotransplantation, vol. 11, no. 5, pp. 444-451. https://doi.org/10.1111/j.1399-3089.2004.00163.x

In vivo gene transfer of endo-β-galactosidase C removes αGal antigen on erythrocytes and endothelial cells of the organs. / Miki, Yusuke; Maruyama, Shoichi; Liu, Da Ge; Kobayashi, Takaaki; Sato, Fumihiko; Shimizu, Hideaki; Kato, Sawako; Sato, Waichi; Morita, Yoshiki; Yuzawa, Yukio; Muramatsu, Takashi; Matsuo, Seiichi.

In: Xenotransplantation, Vol. 11, No. 5, 01.09.2004, p. 444-451.

Research output: Contribution to journalArticle

TY - JOUR

T1 - In vivo gene transfer of endo-β-galactosidase C removes αGal antigen on erythrocytes and endothelial cells of the organs

AU - Miki, Yusuke

AU - Maruyama, Shoichi

AU - Liu, Da Ge

AU - Kobayashi, Takaaki

AU - Sato, Fumihiko

AU - Shimizu, Hideaki

AU - Kato, Sawako

AU - Sato, Waichi

AU - Morita, Yoshiki

AU - Yuzawa, Yukio

AU - Muramatsu, Takashi

AU - Matsuo, Seiichi

PY - 2004/9/1

Y1 - 2004/9/1

N2 - Background: The presence of Galα1-3Galβ1-4GlcNAc (αGal) in pigs is a formidable barrier for pig-to-primate xenotransplantation. We have reported that administration of recombinant endo-β-galactosidase C (EndoGalC) removes αGal on porcine erythrocytes and kidneys. The present study examined the effects of EndoGalC gene therapy on αGal suppression. Methods: Naked plasmid DNA encoding Igκ-EndoGalC was given to rats by rapid tail vein injection. The expression of αGal in the heart and kidney were studied by lectin staining followed by computer-assisted quantitative analyses. αGal expression on erythrocytes was determined by flow cytometric analyses. Enzymatic activity of EndoGalC in the serum was also determined by evaluating the capacity of EndoGalC in removing αGal epitopes. Elimination of αGal was further studied by injecting antibodies against αGal to rats 2 days after the gene transfer. Results: Administration of 1 mg of Igκ-EndoGalC/pCAGGS plasmid eliminated αGal from the vascular endothelium of the heart and kidney on day 1 and day 2. Between days 4 and 7, αGal started to reappear, but remained suppressed. No serious adverse effect was observed in rats treated with EndoGalC. Flow cytometric analyses showed that Endo-GalC digested 97% of αGal on erythrocytes when measured 4 days after the gene transfer. Enzymatic activity of EndoGalC in the serum peaked on day 1, and significant levels were still observed on day 7. When antibodies against αGal were given, rats treated with EndoGalC showed no change, while all control rats died within 40 min. Conclusions: The present study demonstrates the potential of an EndoGalC gene transfer, using a hydrodynamics-based delivery system, in eliminating αGal from endothelial cells in vivo. The results also ensured that EndoGalC is not harmful suggesting that the production of pigs overexpressing EndoGalC would be a reasonable alternative to pigs deficient in α1,3galactosyltransferase.

AB - Background: The presence of Galα1-3Galβ1-4GlcNAc (αGal) in pigs is a formidable barrier for pig-to-primate xenotransplantation. We have reported that administration of recombinant endo-β-galactosidase C (EndoGalC) removes αGal on porcine erythrocytes and kidneys. The present study examined the effects of EndoGalC gene therapy on αGal suppression. Methods: Naked plasmid DNA encoding Igκ-EndoGalC was given to rats by rapid tail vein injection. The expression of αGal in the heart and kidney were studied by lectin staining followed by computer-assisted quantitative analyses. αGal expression on erythrocytes was determined by flow cytometric analyses. Enzymatic activity of EndoGalC in the serum was also determined by evaluating the capacity of EndoGalC in removing αGal epitopes. Elimination of αGal was further studied by injecting antibodies against αGal to rats 2 days after the gene transfer. Results: Administration of 1 mg of Igκ-EndoGalC/pCAGGS plasmid eliminated αGal from the vascular endothelium of the heart and kidney on day 1 and day 2. Between days 4 and 7, αGal started to reappear, but remained suppressed. No serious adverse effect was observed in rats treated with EndoGalC. Flow cytometric analyses showed that Endo-GalC digested 97% of αGal on erythrocytes when measured 4 days after the gene transfer. Enzymatic activity of EndoGalC in the serum peaked on day 1, and significant levels were still observed on day 7. When antibodies against αGal were given, rats treated with EndoGalC showed no change, while all control rats died within 40 min. Conclusions: The present study demonstrates the potential of an EndoGalC gene transfer, using a hydrodynamics-based delivery system, in eliminating αGal from endothelial cells in vivo. The results also ensured that EndoGalC is not harmful suggesting that the production of pigs overexpressing EndoGalC would be a reasonable alternative to pigs deficient in α1,3galactosyltransferase.

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