title: Intracoronary adenovirus-mediated transfer of immunosuppressive cytokine genes prolongs allograft survival.
abstract: BACKGROUND
  Intracoronary transfer and expression of recombinant genes in the intact heart is now feasible. In the transplant setting, local modulation of host immune responses by a genetically modified allograft may offer an attractive alternative to systemic immunosuppression.
  
  
  METHODS
  We tested the efficacy and in vivo effect of intracoronary transfer of two immunosuppressive cytokine genes. First-generation E1-deleted adenoviral vectors expressing the Epstein-Barr virus interleukin-10 (AdSvIL10) or human transforming growth factor--beta 1 (AdCMVTGF-beta) were used. Rabbit cardiac allografts were transduced during cold preservation by slow (1 ml/min) intracoronary infusion of 10(10) pfu/gm diluted viral vectors and then implanted heterotopically. Controls included E1-deleted adenovirus (Ad5dI434) and AdCMVLacZ. Beating allografts were collected on day 4 for analysis of gene transfer efficacy and distribution. Additional grafts were used for evaluation of alloreactivity (n = 34).
  
  
  RESULTS
  Mean allograft viral uptake was 81% (up to 91%). Polymerase chain reactions and reverse transcription-polymerase chain reactions confirmed the presence and expression of both genes in the grafts. beta-Galactosidase staining in AdCMVLacZ-infected grafts demonstrated efficient gene expression, which was highest (100%) in subepicardial regions. More homogeneous transmyocardial distribution of the transgene (in 25% to 40% of cells) could be achieved by pulsatile slow delivery. Allograft survival was 6.9 +/- 0.9 days in controls (n = 12), 11.1 +/- 1.7 days in AdCMVTGF-beta-infected grafts (n = 11, p < 10(-4)), and 11.2 +/- 3 days in AdSvIL10-infected grafts (n = 11, p < 10(-4)). Histologic scores (blinded) showed significantly (p < 0.005) higher regression coefficients for rejection in controls compared with both cytokine-transduced groups. Perioperative administration of cyclosporine A (INN: ciclosporin) to recipients had no effect on survival of AdCMVTGF-beta-infected grafts but reduced survival of AdSvIL10-infected grafts.
  
  
  CONCLUSIONS
  Intracoronary gene transfer of immunosuppressive cytokines to cardiac allografts is efficient and effectively prolongs graft survival. Vectors that would induce long-term expression of such genes may make this approach clinically applicable.
authors.affiliation: Division of Cardiothoracic Surgery, UCLA Medical Center 90095, USA.
authors.forename: R
authors.initials: R
authors.lastname: Brauner
authors.forename: M
authors.initials: M
authors.lastname: Nonoyama
authors.forename: H
authors.initials: H
authors.lastname: Laks
authors.forename: D C
authors.initials: DC
authors.lastname: Drinkwater
authors.forename: S
authors.initials: S
authors.lastname: McCaffery
authors.forename: T
authors.initials: T
authors.lastname: Drake
authors.forename: A J
authors.initials: AJ
authors.lastname: Berk
authors.forename: L
authors.initials: L
authors.lastname: Sen
authors.forename: L
authors.initials: L
authors.lastname: Wu
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006084:Graft Rejection / Q000517:prevention & control*; D006085:Graft Survival / Q000276:immunology*; D016027:Heart Transplantation / Q000276:immunology*; D006801:Humans; D007165:Immunosuppression Therapy / Q000379:methods*; D007261:Infusions, Intra-Arterial; D016753:Interleukin-10 / Q000008:administration & dosage / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011817:Rabbits; D016212:Transforming Growth Factor beta / Q000008:administration & dosage / Q000235:genetics*; D016042:Transplantation, Heterotopic; D014184:Transplantation, Homologous; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D016212:Transforming Growth Factor beta; D016753:Interleukin-10; D001616:beta-Galactosidase