title: Design, expression and characterization of a novel coexpression system of two antiarthritic molecules.
abstract: The complexity of rheumatoid arthritis (RA) pathogenesis makes combined blockade of multiple targets an attractive therapeutic strategy. The combination therapy with anti-TNF plus anti-T-cell has been mostly reported to provide greater efficacy than anti-TNF alone. TNFR (p75)-Fc fusion protein, which has been proven effective in clinics, is chosen as the TNF antagonist in this study. CTLA4-FasL fusion molecule, which has been well characterized in our previous studies for its suppressive effect in rat arthritis model, is chosen as the T-cell antagonist. In this study, furin cleavage site and 2A self-processing sequence were introduced to link upstream TNFR-Fc and downstream CTLA4-FasL and mediate separate coexpression of the two fusion proteins in a single recombinant adeno-associated virus (rAAV) vector. Using this expression system, we generated two fusion proteins with same size as their individual counterparts in vitro and in vivo, and the proteins desirably retained their parent biological activities. In vivo results demonstrated that furin-2A technology is able to regulate separate coexpression of these proteins under arthritic inflammatory conditions. This study describes a single rAAV vector for production of two antiarthritic molecules antagonizing both TNF and T cells, which may serve as an attractive expression system for RA gene therapy.
authors: Department of Translational Medicine, Beijing Institute of Basic Medical Sciences, Beijing 100850, China. zhangweiyh@126.com
authors: Wei
authors: W
authors: Zhang
authors: Fang
authors: F
authors: Wang
authors: Jinqi
authors: J
authors: Yan
authors: Xiaojun
authors: X
authors: Zhang
authors: Yu
authors: Y
authors: Wang
authors: Yunbo
authors: Y
authors: Jiang
authors: Lin
authors: L
authors: Wang
authors: Yuanji
authors: Y
authors: Xu
authors: Jiyun
authors: J
authors: Yu
mesh_terms: D000818:Animals; D001172:Arthritis, Rheumatoid / Q000235:genetics / Q000628:therapy* / Q000821:virology; D060908:CTLA-4 Antigen / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D000229:Dependovirus / Q000235:genetics / Q000378:metabolism; D015195:Drug Design; D053222:Fas Ligand Protein / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D005260:Female; D015870:Gene Expression; D015316:Genetic Therapy; D006801:Humans; D051379:Mice; D015202:Protein Engineering; D011499:Protein Processing, Post-Translational; D051381:Rats; D011917:Rats, Inbred Lew; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism
chemical_list: D060908:CTLA-4 Antigen; D053222:Fas Ligand Protein; D047889:Receptors, Tumor Necrosis Factor, Type II; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha