title: Naked DNA transfer of Factor VIII induced transgene-specific, species-independent immune response in hemophilia A mice.
abstract: The development of antibodies to a previously unexpressed protein product may limit the success of human gene therapy approaches. We inserted B-domain-deleted factor VIII (FVIII) cDNA of human, canine, or murine origin into the multiple cloning site of a liver-specific vector, pBS-HCRHPI-A, to yield plasmids pBS-HCRHPI-FVIIIA, pBS-HCRHPI-cFVIIIA, and pBS-HCRHPI-mFVIIIA, respectively. Fifty micrograms of each plasmid in 2 ml of solution was rapidly injected into the tail vein of three groups of hemophilia A mice. Factor VIII levels ranging from 3 to 12 IU/ml were obtained from all three groups (normal is 1 IU/ml in human plasma) 3 days after treatment. These initial very high levels of functional human, canine, or murine factor VIII, however, fell gradually to undetectable levels within 2-3 weeks, and their disappearance correlated with the generation of high-titer, inhibitory anti-FVIII antibodies. Notably, this immune response occurred independent of the species of origin of the exogenous factor VIII. Antibody titers to factor VIII were detected beginning at 2 weeks, reached a plateau and remained at high levels for over 6 months. The majority of anti-hFVIII IgG was IgG1 isotype specific, suggesting a humoral response mediated by Th2-induced signals. Consistent with this idea, in a separate group of mice treated with pBS-HCRHPI-FVIIIA, transient immunosuppression by cyclophosphamide significantly delayed (5/6) or abolished (1/6) inhibitory antibody formation against the transgene.
authors.affiliation: Department of Pediatrics and Department of Medicine, University of Washington, Seattle, WA 98195, USA.
authors.forename: Peiqing
authors.initials: P
authors.lastname: Ye
authors.forename: Arthur R
authors.initials: AR
authors.lastname: Thompson
authors.forename: Rita
authors.initials: R
authors.lastname: Sarkar
authors.forename: Zhenping
authors.initials: Z
authors.lastname: Shen
authors.forename: David P
authors.initials: DP
authors.lastname: Lillicrap
authors.forename: Randal J
authors.initials: RJ
authors.lastname: Kaufman
authors.forename: Hans D
authors.initials: HD
authors.lastname: Ochs
authors.forename: David J
authors.initials: DJ
authors.lastname: Rawlings
authors.forename: Carol H
authors.initials: CH
authors.lastname: Miao
mesh_terms: D000818:Animals; D000917:Antibody Formation / Q000187:drug effects; D003520:Cyclophosphamide / Q000494:pharmacology; D004247:DNA / Q000008:administration & dosage; D004285:Dogs; D005169:Factor VIII / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000032:analysis / Q000235:genetics; D006467:Hemophilia A / Q000276:immunology* / Q000628:therapy; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis / Q000378:metabolism; D008099:Liver / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D010446:Peptide Fragments / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D010957:Plasmids / Q000032:analysis / Q000235:genetics; D019076:Transgenes
chemical_list: C451034:B-domain-deleted factor VIII; D007074:Immunoglobulin G; D010446:Peptide Fragments; D003520:Cyclophosphamide; D005169:Factor VIII; D004247:DNA