title: Anti-arthritic effects of FasL gene transferred intra-articularly by an inducible lentiviral vector containing improved tet-on system.
abstract: The objective of this study is to construct and identify an inducible lentiviral vector containing improved tet-on system and FasL gene and observe its effects on pristane-induced arthritis (PIA). FasL gene was amplified from the spleen of Lewis rats by RT-PCR. The tet-on system was improved with insertion of a chicken chromatin insulator (cHS4) element and an rtTA-dependent, tet-responsive element containing modifications of the tetO sequence (TRE-tight1). Pro-apoptosis effect of the vector pTREFasLcHS4V16 on synovial cells was evaluated by flow cytometer in vitro. Anti-arthritis effects of the vector on PIA after intra-articular injection were observed by clinical evaluation and joint histology. Cytokines in synovial tissue were measured by ELISA. The recombinant inducible lentiviral vector pTREFasLcHS4V16 was successfully constructed. The expression response and the pro-apoptosis effects of the vector were doxycycline dose-dependent. The vector injected intra-articularly attenuated the severity of PIA and decreased the level of cytokines in inflamed joints. pTREFasLcHS4V16 with an improved tet-on system can precisely regulate the expression of FasL gene and apoptosis. Anti-arthritis effects were observed after intra-articular injection of the inducible vector.
authors.affiliation: Department of Anesthesiology, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, 310009, China.
authors.forename: Qingyu
authors.initials: Q
authors.lastname: Shi
authors.forename: Xuebi
authors.initials: X
authors.lastname: Tian
authors.forename: Yinlin
authors.initials: Y
authors.lastname: Zhao
authors.forename: Huiyu
authors.initials: H
authors.lastname: Luo
authors.forename: Yuke
authors.initials: Y
authors.lastname: Tian
authors.forename: Ailin
authors.initials: A
authors.lastname: Luo
mesh_terms: D000818:Animals; D000843:Ankle Joint / Q000378:metabolism* / Q000473:pathology; D017209:Apoptosis; D001169:Arthritis, Experimental / Q000209:etiology / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D004318:Doxycycline / Q000494:pharmacology*; D053222:Fas Ligand Protein / Q000096:biosynthesis* / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D018836:Inflammation Mediators / Q000378:metabolism; D007270:Injections, Intra-Articular; D016086:Lentivirus / Q000235:genetics*; D051381:Rats; D011917:Rats, Inbred Lew; D012720:Severity of Illness Index; D013583:Synovial Membrane / Q000378:metabolism / Q000473:pathology; D013729:Terpenes; D014161:Transduction, Genetic
chemical_list: D016207:Cytokines; D053222:Fas Ligand Protein; C505662:Faslg protein, rat; D018836:Inflammation Mediators; D013729:Terpenes; C009042:pristane; D004318:Doxycycline