RRC ID 4769
Author Yokoo T, Ohashi T, Utsunomiya Y, Shiba H, Shen JS, Hisada Y, Eto Y, Kawamura T, Hosoya T.
Title Inflamed glomeruli-specific gene activation that uses recombinant adenovirus with the Cre/loxP system.
Journal J Am Soc Nephrol
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.
Volume 12(11)
Pages 2330-2337
Published 2001-11-1
DOI 10.1681/ASN.V12112330
PII 12/11/2330
PMID 11675409
MeSH Adenoviridae / genetics Animals Base Sequence / genetics Basement Membrane / immunology Bone Marrow Cells / drug effects Bone Marrow Cells / physiology Bone Marrow Transplantation Cells, Cultured Female Gene Expression / drug effects Gene Expression Regulation* Gene Transfer Techniques* Genes, Switch Glomerulonephritis / genetics* Immune Sera / pharmacology Integrases / genetics Kidney Glomerulus / immunology Lac Operon Lipopolysaccharides / pharmacology Mice Mice, Inbred DBA Promoter Regions, Genetic / physiology Recombination, Genetic Transcriptional Activation Transfection Viral Proteins / genetics
IF 9.274
Times Cited 10
DNA material AxCANCre (RDB01748) AxCANLacZ (RDB01749) AxCALNLNZ (RDB01750).