title: Adeno-associated virus production of soluble tumor necrosis factor receptor neutralizes tumor necrosis factor alpha and reduces arthritis.
abstract: The major limitation of adenovirus is its association with induction of an inflammatory response and relatively short-term production of the gene therapy transgene product. Adeno-associated virus (AAV) is a 4.68-kb single-strand DNA virus that contains ITRs for viral replication and a packaging signal, and also has been engineered to contain therapeutic genes up to 5 kb in length. Transduction of recombinant AAV (rAAV) results in low inflammatory response and long-term expression. We have cloned a low-immunogenic form of human sTNFRI (sTNFRI2.6D) into AAV (rAAVsTNFRI). This vector was analyzed for its ability to transfect and neutralize the effect of TNF-alpha on primary rheumatoid arthritis synovial fibroblast (RASFs). The rAAVsTNFRI was transduced into the cells at 1.8 x 10(1), 1.8 x 10(2), and 1.8 x 10(3) viral particles per cell. There was greater than 90% neutralization of TNF-alpha at 1.8 x 10(3) viral particles/cell. There was a significant decrease in the synovial cell hyperplasia and cartilage and bone destruction in human TNF-alpha transgenic mice treated intraarticularly with rAAVsTNFRI. These results indicate that the low-immunogenic and long-term expressing vector, rAAVsTNFRI, can be used to deliver the soluble TNF-alpha in vitro and in vivo and effectively reduce the severity of arthritis.
authors.affiliation: Division of Clinical Immunology and Rheumatology, Department of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors.forename: H G
authors.initials: HG
authors.lastname: Zhang
authors.forename: J
authors.initials: J
authors.lastname: Xie
authors.forename: P
authors.initials: P
authors.lastname: Yang
authors.forename: Y
authors.initials: Y
authors.lastname: Wang
authors.forename: L
authors.initials: L
authors.lastname: Xu
authors.forename: D
authors.initials: D
authors.lastname: Liu
authors.forename: H C
authors.initials: HC
authors.lastname: Hsu
authors.forename: T
authors.initials: T
authors.lastname: Zhou
authors.forename: C K
authors.initials: CK
authors.lastname: Edwards
authors.forename: J D
authors.initials: JD
authors.lastname: Mountz
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics* / Q000378:metabolism; D001168:Arthritis / Q000473:pathology / Q000628:therapy*; D001169:Arthritis, Experimental / Q000628:therapy; D001172:Arthritis, Rheumatoid / Q000473:pathology*; D002478:Cells, Cultured; D017364:Collagenases / Q000187:drug effects / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics*; D005260:Female; D005347:Fibroblasts / Q000821:virology; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D007739:L Cells; D051379:Mice; D008822:Mice, Transgenic; D009132:Muscles / Q000821:virology; D018124:Receptors, Tumor Necrosis Factor / Q000235:genetics* / Q000378:metabolism; D047888:Receptors, Tumor Necrosis Factor, Type I; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013585:Synovitis / Q000473:pathology / Q000628:therapy; D018675:Toxicity Tests; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
chemical_list: D015703:Antigens, CD; D018124:Receptors, Tumor Necrosis Factor; D047888:Receptors, Tumor Necrosis Factor, Type I; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D017364:Collagenases