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- Publisher
- Wiley
- Copyright
- © 1997 Munksgaard
- ISSN
- 0908-665X
- eISSN
- 1399-3089
- DOI
- 10.1111/j.1399-3089.1997.tb00185.x
- Publisher site
- See Article on Publisher Site
Abstract
Abstract: The expression of human regulators of complement activation in endothelium of a transplanted organ is an approach to overcoming the complement (C)‐mediated rejection of a xenograft. We designed a replication‐deficient adenovirus (Ad) containing the human CD59 cDNA (AdCD59) in order to induce expression of human CD59 on xenogeneic endothelial cells (EC) and confer protection against human C‐mediated lysis. In vitro transduction of rat EC with AdCD59 led to consistent levels of CD59 expression in all cells and significantly reduced EC lysis induced by human xenogeneic natural antibodies (XNA) binding and C activation. In addition, we analyzed whether Ad transduction had modified the phenotype of EC and the xenogeneic recognition of EC by human serum. For this, we first demonstrated that transduction of rat EC with AdCD59 or an Ad carrying the lacZ gene (AdlacZ) markedly enhanced the expression of some cell‐membrane markers of EC activation, including class I MHC antigens and rat CD59, to levels comparable to those obtained with TNFα Up‐regulation of class I MHC antigens was observed for both mRNA and protein expression. In contrast, AdCD59‐mediated gene transfer slightly increase intercellular adhesion molecule‐1 (ICAM‐1) and did not modify the expression of Crry, a rat complement regulatory molecule. Second, we showed that these phenotypic modifications of EC did not affect the expression of the Galαl‐3Gal epitope and the binding of human XNA. In addition, neither AdlacZ‐mediated transduction, nor TNFα treatment, modified the sensitivity of xenogeneic EC to human serum‐mediated lysis. In conclusion, this study demonstrated that transduction of human CD59 with an adenoviral vector conferred resistance to xenogeneic cultured EC against human C‐mediated lysis but is associated with cellular activation of transduced EC. This finding may have important implications for the in vivo protocols and strategies intended to generate safer Ad vectors.
Journal
Xenotransplantation – Wiley
Published: Nov 1, 1997