Prevention by anti-LFA-1 of acute myoblast death following transplantation.
Myoblast transplantation is a potential treatment for Duchenne muscular dystrophy. One of the problems possibly responsible for the limited success of clinical trials is the rapid death of the myoblasts after transplantation. To investigate this problem, myoblasts expressing beta-galactosidase were injected in the tibialis anterior muscles of mice. Beta-galactosidase activity was reduced by 74.7% after 3 days. Myoblast death observed at 3 days was reduced to 57.2% when the hosts were irradiated. This result suggested that host cells were contributing to this phenomenon. Transplantation in SCID and FK506-treated mice did not reduce cell death, indicating that mortality was not due to an acute specific reaction. In contrast, administration of the anti-LFA-1 (TIB-213) mAb markedly reduced myoblast death at 3 days without altering leukocyte tissue infiltration. We postulated that neutrophils were mediating myoblast mortality by an LFA-1-dependent mechanism. To test this hypothesis, IL-1beta-activated myoblasts were loaded with 6-carboxy-2',7'-dichlorodihydrofluorescein diacetate, di(acetoxymethylester) (DCFH), a marker for oxidative stress. Addition of neutrophils and zymosan-activated serum resulted in a time-dependent DCFH fluorescence; this neutrophil-induced oxidation was considerably inhibited by TIB-213. These results indicate that an effective control of the inflammatory reaction will be necessary for any new clinical trials of myoblast transplantation and suggest that neutrophil-mediated myoblast injury occurs by an LFA-1-dependent pathway.
Guérette
B
B
Université Laval, Département d'Anatomie et Laboratoire de Neurobiologie, Hôpital de l'Enfant-Jésus, Québec, Canada.
Skuk
D
D
Célestin
F
F
Huard
C
C
Tardif
F
F
Asselin
I
I
Roy
B
B
Goulet
M
M
Roy
R
R
Entman
M
M
Tremblay
J P
JP
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000627:therapeutic use; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000627:therapeutic use; D000911:Antibodies, Monoclonal / Q000494:pharmacology* / Q000627:therapeutic use; D017209:Apoptosis; D015415:Biomarkers; D002448:Cell Adhesion; D016923:Cell Death; D002461:Cell Line, Transformed; D005452:Fluoresceins; D017930:Genes, Reporter; D006084:Graft Rejection / Q000473:pathology* / Q000517:prevention & control; D007166:Immunosuppressive Agents / Q000627:therapeutic use; D007249:Inflammation; D007375:Interleukin-1 / Q000494:pharmacology; D016169:Lymphocyte Function-Associated Antigen-1 / Q000276:immunology / Q000502:physiology*; D016177:Macrophage-1 Antigen / Q000502:physiology; D008775:Methylprednisolone / Q000627:therapeutic use; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018101:Mice, Inbred mdx; D016513:Mice, SCID; D018482:Muscle, Skeletal / Q000166:cytology*; D009137:Muscular Dystrophy, Animal / Q000473:pathology / Q000628:therapy*; D009288:Naproxen / Q000627:therapeutic use; D009336:Necrosis; D009504:Neutrophils / Q000276:immunology / Q000473:pathology; D018384:Oxidative Stress; D009195:Peroxidase / Q000032:analysis; D010894:Piroxicam / Q000627:therapeutic use; D011828:Radiation Chimera; D033581:Stem Cell Transplantation; D016559:Tacrolimus / Q000627:therapeutic use; D014916:Whole-Body Irradiation; D015054:Zymosan / Q000494:pharmacology; D001616:beta-Galactosidase / Q000032:analysis
C096548:6-carboxy-2',7'-dichlorodihydrofluorescein diacetate bis(acetoxymethyl) ester; D000893:Anti-Inflammatory Agents; D000894:Anti-Inflammatory Agents, Non-Steroidal; D000911:Antibodies, Monoclonal; D015415:Biomarkers; D005452:Fluoresceins; D007166:Immunosuppressive Agents; D007375:Interleukin-1; D016169:Lymphocyte Function-Associated Antigen-1; D016177:Macrophage-1 Antigen; D010894:Piroxicam; D009288:Naproxen; D015054:Zymosan; D009195:Peroxidase; D001616:beta-Galactosidase; D016559:Tacrolimus; D008775:Methylprednisolone