RRC ID |
4769
|
著者 |
Yokoo T, Ohashi T, Utsunomiya Y, Shiba H, Shen JS, Hisada Y, Eto Y, Kawamura T, Hosoya T.
|
タイトル |
Inflamed glomeruli-specific gene activation that uses recombinant adenovirus with the Cre/loxP system.
|
ジャーナル |
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.
|
巻・号 |
12(11)
|
ページ |
2330-2337
|
公開日 |
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
|
引用数 |
10
|
WOS 分野
|
UROLOGY & NEPHROLOGY
|
リソース情報 |
遺伝子材料 |
AxCANCre (RDB01748)
AxCANLacZ (RDB01749)
AxCALNLNZ (RDB01750). |