Local Gene Delivery and Methods to Control Immune Responses in Muscles of Normal and Dystrophic Dogs
Adeno-associated viral vector (AAV)-mediated gene transfer represents a promising gene replacement strategy for treating Duchenne muscular dystrophy (DMD). However, recent studies demonstrated cellular immunity specific to AAV capsid proteins in animal models, which resulted in liver toxicity and elimination of transgene expression in a human trial of hemophilia B. We have recently developed immunosuppressive strategies to prevent such immunity for successful long-term transgene expression in dog muscle. Here, we describe in detail the immunosuppressive regimens employed in both normal and DMD dogs and provide methods for evaluating the efficiency of the regimens following intramuscular injection of AAV in dogs.
Key wordsAdeno-associated virus AAV Duchenne muscular dystrophy DMD Cellular immunity Muscle Dog cxmd Immunosuppression CSP MMF ATG
We thank Dr. Jeffery Chamberlain for providing vectors used in the protocols and Dr. Kathy High for providing the c-FIX plasmid. We thank E. Zellmer, Diana Jensen, Jenee O’Brian, and E. Finn for technical assistance, and A. Joslyn, B. Steinmetz, and their teams, and M. Spector, DVM, and J Duncan, DVM, for their care of the dogs. We further thank S. Carbonneau, H. Crawford, B. Larson, K. Carbonneau, Vermeulen, and D. Gayle for administrative assistance and manuscript preparation. This work was supported by NIH U54-HD47175 and the Seattle Senator Paul D. Wellstone Muscular Dystrophy Cooperative Research Center, NIH CA15704, NIH AR056949, and by Career Development Award for Z. Wang from the Muscular Dystrophy Association (MDA 114979).
- 26.Storb R, Gluckman E, Thomas ED, Buckner CD, Clift RA, Fefer A, et al. (1974) Treatment of established human graft-versus-host disease by antithymocyte globulin. Blood 44, 57–75.Google Scholar