A novel strategy for preventing human CD8+ cytotoxic T lymphocyte-mediated cytotoxicity against pig endothelial cells by overexpression of pig cellular FLICE-like inhibitory protein (c-FLIP) gene

Transplant Proc. 2006 Dec;38(10):3315-7. doi: 10.1016/j.transproceed.2006.10.082.

Abstract

Human CD8+ cytotoxic T lymphocyte (CTL)-mediated cytotoxicity in xenograft recipients is an important obstacle for successful xenotransplantation of pig organs into humans. In our previous study, we demonstrated that xenocytotoxicity of human CD8+ CTL detrimental to pig endothelial cells (PEC) is mediated mainly by the Fas/FasL apoptotic pathway. Furthermore, we developed new methods to prevent this CTL killing by extracellular remodeling using overexpression of human decoy Fas antigen and membrane-bound human FasL on pig xenograft cells. The cellular FLICE-inhibitory protein (c-FLIP), a caspase-8 inhibitor that lacks the cysteine domain, is a negative regulator of death receptor-mediated apoptosis. c-FLIP proteins exist as long (c-FLIP(L)) and short (c-FLIPs) splice variants, both capable of protecting cells from death receptor-mediated apoptosis. In this report, we have demonstrated that both pig c-FLIPs and pig c-FLIP(L) significantly inhibit human CD8+ CTL-mediated xenocytotoxicity toward stably transfected PEC, although the expression level of pig Fas antigen on cell surface was not changed. These data suggested that intracellular remodeling with overexpression of pig c-FLIP in xenograft cells may decrease the innate cellular responses against xenografts, facilitating long-term xenograft survival.

MeSH terms

  • Animals
  • CASP8 and FADD-Like Apoptosis Regulating Protein / genetics*
  • Cell Line
  • Cytotoxicity, Immunologic*
  • DNA Primers
  • DNA, Complementary
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / immunology*
  • Endothelium, Vascular / physiology
  • Gene Expression Regulation / immunology
  • Graft Survival
  • Humans
  • RNA, Messenger / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Swine
  • T-Lymphocytes, Cytotoxic / immunology*
  • Transfection
  • Transplantation, Heterologous / immunology

Substances

  • CASP8 and FADD-Like Apoptosis Regulating Protein
  • DNA Primers
  • DNA, Complementary
  • RNA, Messenger