Human immune reactivity against liver sinusoidal endothelial cells from GalTα(1,3)GalT-deficient pigs

Daan Van Poll, Yakoov Nahmias, Alejandro Soto-Gutierrez, Mitra Ghasemi, Hiroshi Yagi, Naoya Kobayashi, Martin L. Yarmush, Martin Hertl

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Elimination of galactose-α(1,3)galactose (Gal) expression in pig organs has been previously shown to prevent hyperacute xenograft rejection. However, naturally present antibodies to non-Gal epitopes activate endothelial cells, leading to acute humoral xenograft rejection. Still, it is unknown whether xenogeneic pig liver sinusoidal endothelial cells (LSECs) from α(1,3)galactosyltransferase (GalT)-deficient pigs are damaged by antibody and complement-mediated mechanisms. The present study examined the xeno-antibody response of LSECs from GalT-deficient and wild pigs. Isolated LSEC from wild-type and GalT pigs were expose to human and baboon sera; IgM and IgG binding was analyzed by flow cytometry. Complement activation (C3a and CH50) was quantified in vitro from serum-exposed LSEC cultures using Enzyme-Linked Immuno-Sorbent assay (ELISA). Levels of complement-activated cytotoxicity (CAC) were also determined by a fluorescent Live-Dead Assay and by the quantification of LDH release. IgM binding to GalT knockout (KO) LSECs was significantly lower (80% human and 87% baboon) compare to wild-type pig LSEC. IgG binding was low in all groups. Moreover, complement activation (C3a and CH50) levels released following exposure to human or baboon sera were importantly reduced (42% human and 52% baboon), CAC in GalT KO LSECs was reduced by 60% in human serum and by 72% in baboon serum when compared to wildtype LSECs, and LDH release levels were reduced by 37% and 57%, respectively. LSECs from GalT KO pigs exhibit a significant protection to humoral-induced cell damage compared to LSECs from wild pigs when exposed to human serum. Although insufficient to inhibit xenogeneic reactivity completely, transgenic GalT KO expression on pig livers might contribute to a successful application of clinical xenotransplantation in combination with other protective strategies.

Original languageEnglish
Pages (from-to)783-789
Number of pages7
JournalCell Transplantation
Volume19
Issue number6-7
DOIs
Publication statusPublished - 2010
Externally publishedYes

Fingerprint

Galactosyltransferases
Endothelial cells
Liver
Swine
Endothelial Cells
Papio
Complement C3a
Serum
Antibodies
Complement Activation
Cytotoxicity
Galactose
Heterografts
Immunoglobulin M
Assays
Immunoglobulin G
Chemical activation
Epitopes
Heterologous Transplantation
Flow cytometry

Keywords

  • GalTα(1,3)GalT-knockout pigs
  • Liver endothelial cells
  • Xenotransplantation

ASJC Scopus subject areas

  • Cell Biology
  • Transplantation
  • Biomedical Engineering

Cite this

Van Poll, D., Nahmias, Y., Soto-Gutierrez, A., Ghasemi, M., Yagi, H., Kobayashi, N., ... Hertl, M. (2010). Human immune reactivity against liver sinusoidal endothelial cells from GalTα(1,3)GalT-deficient pigs. Cell Transplantation, 19(6-7), 783-789. https://doi.org/10.3727/096368910X508898

Human immune reactivity against liver sinusoidal endothelial cells from GalTα(1,3)GalT-deficient pigs. / Van Poll, Daan; Nahmias, Yakoov; Soto-Gutierrez, Alejandro; Ghasemi, Mitra; Yagi, Hiroshi; Kobayashi, Naoya; Yarmush, Martin L.; Hertl, Martin.

In: Cell Transplantation, Vol. 19, No. 6-7, 2010, p. 783-789.

Research output: Contribution to journalArticle

Van Poll, D, Nahmias, Y, Soto-Gutierrez, A, Ghasemi, M, Yagi, H, Kobayashi, N, Yarmush, ML & Hertl, M 2010, 'Human immune reactivity against liver sinusoidal endothelial cells from GalTα(1,3)GalT-deficient pigs', Cell Transplantation, vol. 19, no. 6-7, pp. 783-789. https://doi.org/10.3727/096368910X508898
Van Poll, Daan ; Nahmias, Yakoov ; Soto-Gutierrez, Alejandro ; Ghasemi, Mitra ; Yagi, Hiroshi ; Kobayashi, Naoya ; Yarmush, Martin L. ; Hertl, Martin. / Human immune reactivity against liver sinusoidal endothelial cells from GalTα(1,3)GalT-deficient pigs. In: Cell Transplantation. 2010 ; Vol. 19, No. 6-7. pp. 783-789.
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AU - Yagi, Hiroshi

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