title: Inflamed glomeruli-specific gene activation that uses recombinant adenovirus with the Cre/loxP system.
abstract: The authors previously reported that bone marrow-derived CD11b(+)CD18(+) cells could be used as a vehicle to deliver foreign genes into inflamed glomeruli and that this vehicle cell (v-cell) could retard the progression of nephritis by delivering anti-inflammatory molecules. As a next step, the authors tried to establish a switching system by which v-cells are activated only at the inflamed glomeruli. A recombinant adenovirus (Ad) that expressed Cre recombinase under the control of the interleukin-1 beta (IL-1 beta) promoter (AxIL-1pr/Cre) was constructed and transfected into v-cells. After confirming that AxIL-1pr/Cre expresses Cre by lipopolysaccharide (LPS) treatment, AxIL-1pr/Cre was infected together with another Ad bearing a switching reporter unit in which the LacZ gene is activated under the control of the CAG promoter by the Cremediated excisional deletion of interposed stuffer DNA. Only a negligible number of double-infected (Cre/loxPCAG) cells expressed LacZ. This number, however, was significantly increased by LPS, which suggests that LPS-induced Cre effectively deletes the stuffer DNA, which allows for a complete CAG promoter. DBA/2j mice were then transplanted with Cre/loxPCAG cells via a tail vein and treated with anti-glomerular basement membrane (GBM) serum. To trace the transplanted cells, marker v-cells, infected with AxCANLacZ to constitutively express the LacZ gene, were also used. Although transplanted cells expressing LacZ collected in the spleen independent of anti-GBM treatment, they did not express the LacZ gene in the mice transplanted with Cre/loxPCAG cells. On the other hand, transplanted cells were recruited in the glomeruli and expressed the LacZ gene upon anti-GBM treatment. These results suggested that only the v-cells recruited in the glomeruli could be switched on and activate foreign genes.
authors.affiliation: Division of Nephrology and Hypertension, Department of Internal Medicine, Jikei University School of Medicine, Tokyo, Japan.|Department of Gene Therapy, Institute of DNA Medicine, Jikei University School of Medicine, Tokyo, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Yokoo
authors.affiliation: Department of Gene Therapy, Institute of DNA Medicine, Jikei University School of Medicine, Tokyo, Japan.|Department of Pediatrics, Jikei University School of Medicine, Tokyo, Japan.
authors.forename: Toya
authors.initials: T
authors.lastname: Ohashi
authors.affiliation: Division of Nephrology and Hypertension, Department of Internal Medicine, Jikei University School of Medicine, Tokyo, Japan.
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Utsunomiya
authors.affiliation: Department of Gene Therapy, Institute of DNA Medicine, Jikei University School of Medicine, Tokyo, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Shiba
authors.affiliation: Department of Gene Therapy, Institute of DNA Medicine, Jikei University School of Medicine, Tokyo, Japan.
authors.forename: Jin Song
authors.initials: JS
authors.lastname: Shen
authors.affiliation: Discovery Research Laboratory, Tanabe Seiyaku Co., Ltd, Osaka, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Hisada
authors.affiliation: Department of Gene Therapy, Institute of DNA Medicine, Jikei University School of Medicine, Tokyo, Japan.|Department of Pediatrics, Jikei University School of Medicine, Tokyo, Japan.
authors.forename: Yoshikatsu
authors.initials: Y
authors.lastname: Eto
authors.affiliation: Division of Nephrology and Hypertension, Department of Internal Medicine, Jikei University School of Medicine, Tokyo, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Kawamura
authors.affiliation: Division of Nephrology and Hypertension, Department of Internal Medicine, Jikei University School of Medicine, Tokyo, Japan.
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Hosoya
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001483:Base Sequence / Q000235:genetics; D001485:Basement Membrane / Q000276:immunology; D001854:Bone Marrow Cells / Q000187:drug effects / Q000502:physiology; D016026:Bone Marrow Transplantation; D002478:Cells, Cultured; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D005812:Genes, Switch; D005921:Glomerulonephritis / Q000235:genetics*; D007106:Immune Sera / Q000494:pharmacology; D019426:Integrases / Q000235:genetics; D007678:Kidney Glomerulus / Q000276:immunology; D007763:Lac Operon; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D008811:Mice, Inbred DBA; D011401:Promoter Regions, Genetic / Q000502:physiology; D011995:Recombination, Genetic; D015533:Transcriptional Activation; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics
chemical_list: D007106:Immune Sera; D008070:Lipopolysaccharides; D014764:Viral Proteins; C045073:Cre recombinase; D019426:Integrases