Myoblast transplantation in non-dystrophic dog.
Dog myoblasts obtained from muscle biopsies were infected in vitro with a defective retroviral vector containing a cytoplasmic beta-galactosidase (beta-Gal) gene. These myoblasts were initially transplanted in the irradiated muscles of SCID mice and beta-Gal positive muscle fibers were observed. beta-Gal myoblasts were also transplanted back either in the donor dogs (autotransplantation model) or in unrelated recipient dogs (allotransplantation model). Following these myoblast injections, a rapid inflammatory reaction developed within the muscle as indicated by an expression of P-selectin and of pro-inflammatory cytokine mRNAs (interleukin 6 (IL-6) and transforming growth factor beta (TGF-beta), and by a neutrophil infiltration. Following either auto- or allotransplantation in inadequately or non-immunosuppressed dogs, a specific immune reaction also developed within 2 weeks as indicated by the infiltration of CD4+ and of CD8+ lymphocytes, the increased expression of IL-10 and granzyme B mRNAs and the presence of antibodies reacting with the injected cells. Some dogs were immunosuppressed with several combinations of FK506, cyclosporine (CsA) and RS-61443. In dogs immunosuppressed with CsA combined with RS-61443, only a few myoblasts and myotubes expressing beta-Gal were observed 1-2 weeks after the transplantation, but no muscle fibers expressing beta-Gal were observed after 4 weeks, and antibodies against the injected cells were formed. In dogs immunosuppressed with FK506 alone, although no antibodies against the injected cells were produced, there were no small cells and no muscle fibers expressing beta-Gal 1 month after the transplantation. However, FK506 triggered diarrhea and vomiting in dogs. When the dogs were immunosuppressed with FK506 combined with CsA and RS-61443, muscle fibers expressing beta-Gal were present 4 weeks after the transplantation and no antibodies reacting with donor myoblasts were detected. These results indicate that the combination of three immunosuppressive agents (i.e., FK506, CsA and RS-61443) is effective in controlling the specific immune reactions following myoblast transplantation in dogs and they underline that the outcome of myoblast transplantation is dependent in part on an adequate immunosuppression. These results obtained here in normal dogs may justify myoblast transplantation in dystrophic dogs despite the side effects of FK506.
Ito
H
H
Département d'Anatomie, Université Laval, Hôpital de l'Enfant-Jésus, Québec, Canada.
Vilquin
J T
JT
Skuk
D
D
Roy
B
B
Goulet
M
M
Lille
S
S
Dugré
F J
FJ
Asselin
I
I
Roy
R
R
Fardeau
M
M
Tremblay
J P
JP
D000818:Animals; D001706:Biopsy; D017690:Cell Transplantation; D002478:Cells, Cultured; D016572:Cyclosporine / Q000627:therapeutic use; D004285:Dogs; D004359:Drug Therapy, Combination; D007166:Immunosuppressive Agents / Q000627:therapeutic use*; D051379:Mice; D016513:Mice, SCID; D009132:Muscles / Q000166:cytology / Q000276:immunology / Q000637:transplantation*; D009137:Muscular Dystrophy, Animal / Q000601:surgery; D009173:Mycophenolic Acid / Q000031:analogs & derivatives / Q000627:therapeutic use; D016559:Tacrolimus / Q000627:therapeutic use; D014183:Transplantation, Heterologous; D014184:Transplantation, Homologous; D001616:beta-Galactosidase / Q000096:biosynthesis
D007166:Immunosuppressive Agents; D016572:Cyclosporine; D001616:beta-Galactosidase; D009173:Mycophenolic Acid; D016559:Tacrolimus