Abstract
We previously developed the "immunogene" approach toward cancer gene therapy using epidermal growth factor receptor (EGFR)-mediated endocytosis. Here, we describe an improved immunogene system, in which the antigen-binding (Fab) fragments of the monoclonal antibody (Ab) B4G7 against the human EGFR were conjugated with poly-L-lysine to form a gene delivery vehicle (designated Fab "immunoporter"). Within 12 hours, the β-galactosidase (β-gal) gene was transferred via the Fab immunoporter to virtually all of the nuclei of human squamous carcinoma A431 cells that overproduce the EGFR, and the β-gal enzyme activity was detected within 24 hours and retained for more than 3 days. The β-gal gene was not transferred into human and mouse cells that were deficient in EGFRs, but it was delivered if those mouse cells were transformed with human EGFR genes. β-gal gene transfer via the Fab immunoporter was inhibited by pretreatment with excess amounts of the Fab fragment. The transfer efficiency of the β-gal gene to A431 cells via the Fab immunoporter was ∼2%, which is as high as the lipofection method and 20- to 100-fold higher than the whole Ab immunoporter. The transfer of the herpes simplex virus thymidine kinase gene into A431 tumor cells as a form of the thymidine kinase/Fab immunogene was successful, and subsequent treatment with ganciclovir induced remarkable suicide effects which conferred 1000-fold higher drug sensitivity. Thus, the Fab immunogene was substantially improved with regard to the whole Ab immunogene and could be used as a potent gene transfer vehicle for the in vivo targeting of EGFR-hyperproducing tumor cells.
| Original language | English |
|---|---|
| Pages (from-to) | 357-364 |
| Number of pages | 8 |
| Journal | Cancer Gene Therapy |
| Volume | 5 |
| Issue number | 6 |
| Publication status | Published - 1998 |
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Keywords
- Anti-epidermal growth factor receptor antibody
- Endocytosis
- Fab
- Immunogene
- Suicide gene
ASJC Scopus subject areas
- Cancer Research
- Genetics
Cite this
Receptor-mediated gene delivery using the Fab fragments of anti-epidermal growth factor receptor antibodies : Improved immunogene approach. / Chen, Jiabing; Gamou, Shinobu; Takayanagi, Atsushi; Ohtake, Yuichiro; Ohtsubo, Masafumi; Shimizu, Nobuyoshi.
In: Cancer Gene Therapy, Vol. 5, No. 6, 1998, p. 357-364.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Receptor-mediated gene delivery using the Fab fragments of anti-epidermal growth factor receptor antibodies
T2 - Improved immunogene approach
AU - Chen, Jiabing
AU - Gamou, Shinobu
AU - Takayanagi, Atsushi
AU - Ohtake, Yuichiro
AU - Ohtsubo, Masafumi
AU - Shimizu, Nobuyoshi
PY - 1998
Y1 - 1998
N2 - We previously developed the "immunogene" approach toward cancer gene therapy using epidermal growth factor receptor (EGFR)-mediated endocytosis. Here, we describe an improved immunogene system, in which the antigen-binding (Fab) fragments of the monoclonal antibody (Ab) B4G7 against the human EGFR were conjugated with poly-L-lysine to form a gene delivery vehicle (designated Fab "immunoporter"). Within 12 hours, the β-galactosidase (β-gal) gene was transferred via the Fab immunoporter to virtually all of the nuclei of human squamous carcinoma A431 cells that overproduce the EGFR, and the β-gal enzyme activity was detected within 24 hours and retained for more than 3 days. The β-gal gene was not transferred into human and mouse cells that were deficient in EGFRs, but it was delivered if those mouse cells were transformed with human EGFR genes. β-gal gene transfer via the Fab immunoporter was inhibited by pretreatment with excess amounts of the Fab fragment. The transfer efficiency of the β-gal gene to A431 cells via the Fab immunoporter was ∼2%, which is as high as the lipofection method and 20- to 100-fold higher than the whole Ab immunoporter. The transfer of the herpes simplex virus thymidine kinase gene into A431 tumor cells as a form of the thymidine kinase/Fab immunogene was successful, and subsequent treatment with ganciclovir induced remarkable suicide effects which conferred 1000-fold higher drug sensitivity. Thus, the Fab immunogene was substantially improved with regard to the whole Ab immunogene and could be used as a potent gene transfer vehicle for the in vivo targeting of EGFR-hyperproducing tumor cells.
AB - We previously developed the "immunogene" approach toward cancer gene therapy using epidermal growth factor receptor (EGFR)-mediated endocytosis. Here, we describe an improved immunogene system, in which the antigen-binding (Fab) fragments of the monoclonal antibody (Ab) B4G7 against the human EGFR were conjugated with poly-L-lysine to form a gene delivery vehicle (designated Fab "immunoporter"). Within 12 hours, the β-galactosidase (β-gal) gene was transferred via the Fab immunoporter to virtually all of the nuclei of human squamous carcinoma A431 cells that overproduce the EGFR, and the β-gal enzyme activity was detected within 24 hours and retained for more than 3 days. The β-gal gene was not transferred into human and mouse cells that were deficient in EGFRs, but it was delivered if those mouse cells were transformed with human EGFR genes. β-gal gene transfer via the Fab immunoporter was inhibited by pretreatment with excess amounts of the Fab fragment. The transfer efficiency of the β-gal gene to A431 cells via the Fab immunoporter was ∼2%, which is as high as the lipofection method and 20- to 100-fold higher than the whole Ab immunoporter. The transfer of the herpes simplex virus thymidine kinase gene into A431 tumor cells as a form of the thymidine kinase/Fab immunogene was successful, and subsequent treatment with ganciclovir induced remarkable suicide effects which conferred 1000-fold higher drug sensitivity. Thus, the Fab immunogene was substantially improved with regard to the whole Ab immunogene and could be used as a potent gene transfer vehicle for the in vivo targeting of EGFR-hyperproducing tumor cells.
KW - Anti-epidermal growth factor receptor antibody
KW - Endocytosis
KW - Fab
KW - Immunogene
KW - Suicide gene
UR - http://www.scopus.com/inward/record.url?scp=0032199731&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032199731&partnerID=8YFLogxK
M3 - Article
C2 - 9917090
AN - SCOPUS:0032199731
VL - 5
SP - 357
EP - 364
JO - Cancer Gene Therapy
JF - Cancer Gene Therapy
SN - 0929-1903
IS - 6
ER -