title: Local high-capacity adenovirus-mediated mCTLA4Ig and mCD40Ig expression prolongs recombinant gene expression in skeletal muscle.
abstract: Multiple forms of muscular dystrophy are due to the absence of cytoskeletal muscle proteins that normally protect the integrity of muscle cells. The lack of any adequate treatments for these devastating diseases propels research toward the development of strategies for gene delivery to skeletal muscle. High-capacity adenoviral vectors (HC-AdV) devoid of all viral coding sequences have been developed to avoid expression of viral proteins by the gene therapy vector. However, the capsid proteins that are an essential component of the input viral vector and any residual helper virus in the vector preparation could induce an immune response. Furthermore, the therapeutic protein provided by a gene transfer vector presents the potential to induce an immune response in a patient who does not express a normal cellular protein due to genetic mutation. Therefore, we hypothesize that some immune suppression will be required with therapeutic gene delivery designed for the treatment of patients with inherited muscle diseases. In this study, we constructed and rescued three HC-AdVs expressing murine CTLA4Ig, murine CD40Ig, or both. The backbone vector without a gene insert was rescued as a negative control vector. The production of relevant proteins from each vector was determined in vitro. In vivo function of each of the immunosuppressant vectors was assayed by co-injection with an enhanced green fluorescent protein (EGFP)-expressing first-generation adenoviral vector (AdEGFP) into the tibialis anterior muscle of C57BL/10 mice. Higher levels of muscle EGFP expression were observed in animals receiving an immunosuppressant vector. Furthermore, the production of total anti-AdV and anti-EGFP antibodies was reduced in mice treated with each of the three immunosuppressant vectors. A second intramuscular administration of AdEGFP alone 4 weeks after the initial co-injection was successful in all immunosuppressant vector-treated groups, but not in the negative control vector-treated group. All groups had a high antibody response to adenoviral proteins after the second injection of AdEGFP alone, indicating that the initial co-injection did not tolerize against vector capsid antigens.
authors.affiliation: Department of Neurology, University of Pittsburgh, Pittsburgh, Pennsylvania 15213, USA.
authors.forename: Z L
authors.initials: ZL
authors.lastname: Jiang
authors.forename: D
authors.initials: D
authors.lastname: Reay
authors.forename: F
authors.initials: F
authors.lastname: Kreppel
authors.forename: A
authors.initials: A
authors.lastname: Gambotto
authors.forename: E
authors.initials: E
authors.lastname: Feingold
authors.forename: S
authors.initials: S
authors.lastname: Kochanek
authors.forename: S A
authors.initials: SA
authors.lastname: McCarthy
authors.forename: P R
authors.initials: PR
authors.lastname: Clemens
mesh_terms: D000069594:Abatacept; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D015703:Antigens, CD; D000943:Antigens, Differentiation / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western; D019013:CD40 Antigens / Q000235:genetics* / Q000378:metabolism; D023201:CD40 Ligand; D060908:CTLA-4 Antigen; D016189:Dystrophin / Q000235:genetics; D005434:Flow Cytometry; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D018796:Immunoconjugates; D007074:Immunoglobulin G / Q000276:immunology; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018482:Muscle, Skeletal / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D013601:T-Lymphocytes / Q000276:immunology
chemical_list: D015703:Antigens, CD; D000943:Antigens, Differentiation; D019013:CD40 Antigens; D060908:CTLA-4 Antigen; C556707:Ctla4 protein, mouse; D016189:Dystrophin; D018796:Immunoconjugates; D007074:Immunoglobulin G; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D023201:CD40 Ligand; D049452:Green Fluorescent Proteins; D000069594:Abatacept