Increased expression of the novel molecule OX-2 is involved in prolongation of murine renal allograft survival

Transplantation. 1998 Apr 27;65(8):1106-14. doi: 10.1097/00007890-199804270-00016.

Abstract

Background: Portal venous (p.v.) peritransplant immunization with dendritic cells from bone marrow cultures, along with cyclosporine (10 mg/kg), produces antigen-specific increased renal allograft survival compared with recipients receiving intravenous (i.v.) immunization. Increased survival is associated with altered cytokine production from recipient T cells restimulated with donor antigen. We used a suppressive subtractive hybridization approach to explore a role in the regulation of transplant rejection for other genes differentially expressed after p.v. immunization.

Methods: Subtractive hybridization was performed using tissue from p.v. and i.v. immunized mice and a novel polymerase chain reaction-based approach. A gene-bank search was used to identify the source of the differentially expressed cDNAs. One product, the mouse homologue of rat OX-2, was further analyzed using Western gels and FACS analysis of dendritic cells (NLDC145+) isolated from p.v.-immunized mice.

Results: Eighty cDNA clones were obtained by suppressive subtractive hybridization. Differential expression was confirmed in Northern RNA blots. One clone showed sequence homology to a gene encoding a molecule on rat dendritic cells (MRC OX-2), with homology to genes encoding the costimulatory molecules CD80 (B7-1) and CD86 (B7-2). In p.v.-immunized mice, a monoclonal antibody to the rat OX-2 molecule identified, by Western blot analysis, increased expression of a molecule with molecular weight (43 kDa) analogous to rat MRC-OX-2; labels (by FACS analysis) indentified increased numbers of a population of cells staining with NLDC145; and blocks indentified increased graft survival.

Conclusion: Our data suggest that OX-2 is functionally important in the increased graft survival seen in p.v.-immunized mice receiving renal allografts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • Antigens, CD
  • Antigens, Surface / analysis
  • Antigens, Surface / biosynthesis*
  • Antigens, Surface / genetics*
  • Base Sequence
  • Bone Marrow Cells / cytology
  • Bone Marrow Cells / immunology
  • Cloning, Molecular
  • Cytokines / biosynthesis
  • DNA Primers
  • DNA, Complementary
  • Dendritic Cells / immunology*
  • Dendritic Cells / transplantation*
  • Graft Survival / immunology*
  • Humans
  • Immunosuppression / methods
  • In Situ Hybridization
  • Isoantigens / immunology
  • Kidney Transplantation / immunology*
  • Membrane Glycoproteins / biosynthesis
  • Mice
  • Mice, Inbred C3H
  • Mice, Inbred C57BL
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Portal Vein / immunology*
  • Rats
  • Sequence Alignment
  • Sequence Homology, Nucleic Acid
  • T-Lymphocytes / immunology
  • Time Factors
  • Transplantation, Homologous

Substances

  • Antibodies, Monoclonal
  • Antigens, CD
  • Antigens, Surface
  • Cytokines
  • DNA Primers
  • DNA, Complementary
  • Isoantigens
  • Membrane Glycoproteins
  • antigens, CD200