title: Induction of immune tolerance to FIX following muscular AAV gene transfer is AAV-dose/FIX-level dependent.
abstract: Direct intramuscular injection (IM) of adeno-associated virus (AAV) has been proven a safe and potentially efficient procedure for gene therapy of many genetic diseases including hemophilia B. It is, however, contentious whether high antigen level induces tolerance or immunity to coagulation factor IX (FIX) following IM of AAV. We recently reported induction of FIX-specific immune tolerance by IM of AAV serotype one (AAV1) vector in mice. We hypothesize that the expression of high levels of FIX is critical to induction of FIX tolerance. In this study, we investigated the correlation among AAV dose, FIX expression, and tolerance induction. We observed that induction of immune tolerance or immunity to FIX was dependent on the dose of AAV1-human FIX (hFIX) given and the level of FIX antigen expressed in both normal and hemophilia mice. We then defined the minimum AAV1-hFIX dose and the lowest level of FIX needed for FIX tolerance. Different from hepatic AAV-hFIX gene transfer, we found that FIX tolerance induced by IM of AAV1 was not driven by regulatory T cells. These results provided further insight into the mechanism(s) of FIX tolerance, contributing to development of hemophilia gene therapy, and optimization of FIX tolerance induction protocols.
authors.affiliation: Division of Hematology/Oncology, Department of Medicine, Immunology Institute, Mount Sinai School of Medicine, New York, New York 10029, USA.
authors.forename: Meagan E
authors.initials: ME
authors.lastname: Kelly
authors.forename: Jiacai
authors.initials: J
authors.lastname: Zhuo
authors.forename: Arpita S
authors.initials: AS
authors.lastname: Bharadwaj
authors.forename: Hengjun
authors.initials: H
authors.lastname: Chao
mesh_terms: D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology; D049109:Cell Proliferation; D000229:Dependovirus / Q000235:genetics*; D005164:Factor IX / Q000235:genetics* / Q000276:immunology*; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics*; D007108:Immune Tolerance / Q000276:immunology*; D007273:Injections, Intramuscular; D051379:Mice; D008810:Mice, Inbred C57BL; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology
chemical_list: D005164:Factor IX