Inflamed glomeruli-specific gene activation that uses recombinant adenovirus with the Cre/loxP system.
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.
Yokoo
Takashi
T
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.
Ohashi
Toya
T
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.
Utsunomiya
Yasunori
Y
Division of Nephrology and Hypertension, Department of Internal Medicine, Jikei University School of Medicine, Tokyo, Japan.
Shiba
Hiroaki
H
Department of Gene Therapy, Institute of DNA Medicine, Jikei University School of Medicine, Tokyo, Japan.
Shen
Jin Song
JS
Department of Gene Therapy, Institute of DNA Medicine, Jikei University School of Medicine, Tokyo, Japan.
Hisada
Yutaka
Y
Discovery Research Laboratory, Tanabe Seiyaku Co., Ltd, Osaka, Japan.
Eto
Yoshikatsu
Y
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.
Kawamura
Tetsuya
T
Division of Nephrology and Hypertension, Department of Internal Medicine, Jikei University School of Medicine, Tokyo, Japan.
Hosoya
Tatsuo
T
Division of Nephrology and Hypertension, Department of Internal Medicine, Jikei University School of Medicine, Tokyo, Japan.
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
D007106:Immune Sera; D008070:Lipopolysaccharides; D014764:Viral Proteins; C045073:Cre recombinase; D019426:Integrases